4,5,6,7-tetrahydro-1h-imidazo[4,5-c]pyridine and 1,4,5,6,7,8-hexahydroimidazo[4,5-d]azepine derivatives as janus kinase inhibitors

ABSTRACT

There are provided compounds of formula (I), wherein X, Ak, s, L, Q, R 1  and R 2  have meanings given in the description, which compounds have antiinflammatory activity (e.g. through inhibition of one or more of members of the JAK family) and have use in therapy, including in pharmaceutical combinations, especially in the treatment of inflammatory diseases, including inflammatory diseases of the lung, eye and intestines.

This invention relates, inter alia, to compounds which are antiinflammatory agents (e.g., through inhibition of one or more of members of the Janus kinase (JAK) family). The invention also relates to the use of such compounds in therapy, including in mono- and combination therapies, especially in the treatment of inflammatory diseases, including inflammatory diseases of the lung (such as asthma and chronic obstructive pulmonary disease (COPD)), eye (such as uveitis or dry eye disease (DED, also known as keratoconjunctivitis sicca and xerophthalmia)) and gastrointestinal tract (such as Crohn's disease (CD) and ulcerative colitis (UC)).

The listing or discussion of an apparently prior-published document in this specification should not necessarily be taken as an acknowledgement that the document is part of the state of the art or is common general knowledge.

The JAKs are a family of intracellular tyrosine kinases that play an essential role in the signalling of numerous cytokines implicated in the pathogenesis of inflammatory diseases and are critical to both innate and adaptive immunities (Clark, J. D.; Flanagan, M. E.; Telliez, J.-B. J. Med. Chem. 2014, 57, 5023-5038). The family comprises four members, JAK1, JAK2 and TYK2—all of which are expressed ubiquitously—and JAK3, found only in hematopoietic cells. These enzymes show high sequence homology and are constitutively bound to the cytoplasmic tail of cytokine receptors. When a cytokine binds to its receptor, multimerization (dimerization or higher order complexes) of receptor subunits occurs, bringing the JAK enzymes associated with each subunit proximal to one another. JAK family members then auto- and/or trans-phosphorylate, which triggers a series of phosphorylation events resulting ultimately in the phosphorylation and activation of signal transducers and activators of transcription (STAT) proteins. A phosphorylated STAT dimer then translocates to the nucleus of the cell where it binds to target genes to modulate their expression and alter cellular function. Importantly, there are no known compensatory pathways around JAK/STAT signalling and, as such, the JAK enzymes are essential in regulating the cytokines that signal through these pathways. As a result of their critical role in cytokine signalling, the JAK enzymes have become targets for drug discovery and development endeavours that have given rise to two marketed products, tofacitinib and ruxolitinib, as well as various compounds in development (Norman, P. Expert Opin. Investig. Drugs 2014, 23, 1067-1077). Of the two commercialised products, pan-JAK inhibitor tofacitinib (Flanagan, M. E., et al. J. Med. Chem. 2010, 53, 8468-8484) is more relevant for inflammatory diseases, being marketed for the treatment of rheumatoid arthritis (Yamaoka, K.; Tanaka, Y. Expert Opin. Pharmacother. 2014, 15, 103-113) and investigated in the clinic for inflammatory bowel diseases (IBD), such as UC (Sandborn, W. J., et al. New Engl. J. Med. 2012, 367, 616-624; Vuitton, L.; Koch, S.; Peyrin-Biroulet, L. Curr. Drug Targ. 2013, 14, 1385-1391; Panes, J., et al. BMC Gastroenterol. 2015, 15, 14), while JAK1/JAK2 inhibitor ruxolitinib is marketed for the treatment of myelofibrosis (Mesa, R. A.; Yasothan, U.; Kirkpatrick, P. Nat. Rev. Drug Discov. 2012, 11, 103-104). Tofacitinib and other JAK inhibitors have been mooted as potential therapies for other immunological disorders, including COPD (Barnes, P. J. Nat. Rev. Drug Discov. 2013, 12, 543-559; Fenwick, P. S., et al. PLoS ONE 2015, 10(6), e0128757), DED (Beals, C. R.; Woldemussie, E. US Patent Application Publication US 2010/0267751, 21 Oct. 2010; Liew, S. H., et al. Ophthalmology 2012, 119, 1328-1335; Huang, J.-F., et al. Ophthalmology 2012, 119, e43-e50) and uveitis (Huang, J.-F.; Zhang, Y.; Hirakawa, B., 2013 Association for Research in Vision and Ophthalmology Annual Meeting, Seattle, USA, 5-9 May 2013, Abstract 2536).

JAK kinases function as homo or heterodimers dimers which are specific to cytokine receptor subunits. For example, JAK1-JAK3 heterodimers associate with the γ-common chain of receptors to control signalling associated with IL2, IL4, IL7, IL9, IL15 and IL21, cytokines predominantly associated with adaptive immune functions. JAK1, however, also functions as a heterodimer with JAK2 and TYK2 to regulate signalling through a wide array of cytokine receptors. In this manner, JAK1 modulates the signalling of several proinflammatory cytokines associated with the innate immune response, such as IL6 and the type I interferons. JAK2 is the only member of the JAK family that can operate as a homodimer. In this combination, JAK2 controls the signalling of various cytokines and growth factors, such as IL3, IL5, granulocyte macrophage colony-stimulating factor, erythropoietin and thrombopoietin.

The potential importance of JAK1 to IBD is emphasised by the fact that two selective JAK1 inhibitors, filgotinib (GLPG0634) and ABT-494, are in phase 2 clinical trials for CD (Galien, R., et al. Gastroenterology 2014, 146 (Suppl 1), S-49, Abstract 188). Filgotinib is of particular interest for the treatment of IBD, given that, when colon biopsies from IBD patients were treated with this compound, the regulation of IL6 and MX1 expression and a relationship with STAT3 phosphorylation were observed (Dupont, S., et al. Inflamm. Res. 2015, 64 (Suppl 2), S202, Abstract B252). Furthermore, filgotinib has demonstrated efficacy in the mouse dextran sodium sulfate-induced model of colitis, with efficacy being associated with the inhibition of STAT3 phosphorylation in the inflamed colon (Merciris, D., et al. 9th Congress of the European Crohn's and Colitis Organisation, Copenhagen, Denmark, 20-22 Feb. 2014, Abstract P072). JAK2 is implicated in that polymorphisms of the corresponding gene are risk factors for both CD and UC, especially in Caucasians (Zhang, J. X., et al. Inflammation 2014, 37, 793-800). In addition, many components of the IL23 pathway, including JAK2 and TYK2, are true IBD susceptibility genes, suggesting a critical role for this pathway in maintaining intestinal immune homeostasis (Lees, C. W., et al. Gut 2011, 60, 1739-1753; Khor, B.; Gardet, A.; Xavier, R. J. Nature 2011, 474, 307-317). It should also be noted that, when administered orally, the JAK1/2 inhibitor AZD1480 inhibited colitis-associated colon cancer, a model of inflammation-driven tumorigenesis (Stuart, E., et al. Mol. Cancer Ther. 2014, 13, 468-474). This is important, given that patients with UC and CD of the colon have an increased risk of developing colorectal cancer (Farraye, F. A., et al. Gastroenterology 2010, 138, 738-745). The importance of JAK3 in immune homeostasis has been underscored by the observations that loss-of-function mutations in humans result in a severe combined immunodeficiency phenotype (Casanova, J.-L.; Holland, S. M.; Notarangelo, L. D. Immunity 2012, 36, 515-528). Furthermore, the selective JAK3 inhibitor JANEX1 demonstrated activity in preclinical models of colitis (Uckun, F. M., et al. Bioorg. Med. Chem. 2008, 16, 1287-1298). Nonetheless, it has been indicated that JAK3 inhibition alone is not sufficient to achieve maximal anti-inflammatory efficacy and that additional inhibition of JAK1 is necessary to enhance cellular activity (Thoma, G.; Drückes, P.; Zerwes, H.-G. Bioorg. Med. Chem. Lett. 2014, 24, 4617-4621).

As indicated above, all of the JAK enzymes are associated with IBD in one way or another. As such, pan-JAK inhibitors may offer the best chance of achieving efficacy in patients. In this regard, the pan-JAK inhibitor tofacitinib has demonstrated (Boland, B. S.; Sandborn, W. J.; Chang, J. T. Gastroenterol. Clin. N. Am. 2014, 43, 603-617) dose-dependent efficacy in Phase 2 clinical trials for UC, albeit at higher doses than that approved for the treatment of rheumatoid arthritis (5 mg twice-daily). Phase 3 studies are currently underway to evaluate the efficacy of this compound (10 mg twice-daily) as an induction therapy in patients with active UC, and at 5 and 10 mg twice-daily as a maintenance therapy in UC patients who have responded to tofacitinib induction therapy. However, as a result of systemic JAK inhibition, tofacitinib therapy is associated with dose-limiting side effects, in particular, opportunistic infections, dose-related lipid abnormalities and bone marrow suppression, which is mediated through JAK2 inhibition and results in anaemia, thrombocytopenia and neutropenia. Thus, although tofacitinib has demonstrated efficacy in UC patients, there is a need for efficacious agents exhibiting an improved side effect profile.

When administered by inhalation, clinical pan-JAK inhibitor PF06263276 (Reference Compound; Coe, J. W., et al. WO2013/014567, 31 Jan. 2013) has been reported to attenuate IL6-stimulated pSTAT induction in mouse lung. In addition, following topical administration, this compound reduced mouse ear swelling induced by an IL23 injection (Jones, P. Frontiers in Medicinal Chemistry 2015, Antwerp, Belgium, 14-16 Sep. 2015, Abstract IL23). The crystal structure of PF06263276 bound to JAK2 reveals that this compound interacts with the kinase employing the Type 1.5 binding mode (Foloppe, N., et al. Bioorg. Med. Chem. 2006, 14, 1792-1804; Zuccotto, F., et al. J. Med. Chem. 2010, 53, 2681-2694; Wang, T., et al. Bioorg. Med. Chem. Lett. 2010, 20, 153-156) such that its “Northwestern” phenol terminus acts as both a hydrogen bond donor and a hydrogen bond acceptor, binding it deeply within a hydrophobic pocket. The binding mode observed results in PF06263276 exhibiting slow-offset kinetics and an extended cellular duration of action.

We have now discovered, surprisingly, that compounds bearing certain aminoheteroaryl substituents inhibit one or more JAK enzymes and therefore possess good anti-inflammatory properties.

Thus, according to a first aspect of the invention, there is provided a compound of formula I,

wherein: X represents halo; Ak represents C₁₋₆ alkyl optionally substituted by one or more fluorine atoms; s represents 1 or 2; L represents C(O), S(O)₂, CH₂ or a bond; Q represents a 6-membered heteroaromatic ring containing 1, 2 or 3 N atoms; R¹ represents R³; R² represents H, R³, C₁₋₆ alkyl or C₃₋₇ cycloalkyl, which latter two groups are optionally substituted by one or more substituents selected from C₁₋₄ alkyl, halo, hydroxy and oxo; or R¹ and R², together with the N-atom to which they are attached, form a 4- to 7-membered heterocyclic group that is fully saturated or partially unsaturated, which heterocyclic group contains one N atom (the atom to which R¹ and R² are attached) and wherein

-   (a) the heterocyclic group contains one further heteroatom that is S     and is optionally substituted by one or more substituents selected     from halo, hydroxy, oxo, C₁₋₄ alkyl, C₁₋₄ alkoxy and C₁₋₄     hydroxyalkyl or -   (b) the heterocyclic group optionally contains one or more further     heteroatoms selected from N, O and S, is substituted by one or more     substituents selected from C₁₋₄ hydroxyalkyl, C₁₋₄ carboxyalkyl and     —(C₁₋₄ alkylene)-NR^(a)R^(b), and is optionally further substituted     by one or more substituents selected from halo, hydroxy, oxo, CO₂H,     C₁₋₄ alkyl and C₁₋₄ alkoxy or -   (c) the heterocyclic group is fused to a fully saturated or     partially unsaturated 5- or 6-membered heterocyclic ring, which     heterocylic ring contains one or more heteroatoms selected from from     N, O and S and is optionally substituted with one or more     substituents selected from oxo and C₁₋₄ alkyl;     R³ represents     -   Het^(x) or     -   C₁₋₆ alkyl substituted by         -   CO₂H,         -   —(OCH₂CH₂)₀₋₄OR^(c),         -   Het^(y),         -   Het^(z),         -   NR^(d)C(O)R^(e) or         -   S(O)₁₋₂R^(f),     -   which C₁₋₆ alkyl group is optionally further substituted by one         or more substituents selected from halo, hydroxy and oxo;         R^(a), R^(b), R^(c), R^(d) and R^(e) independently represent H         or C₁₋₄ alkyl,         or R^(a) and R^(b), together with the N-atom to which they are         attached, form a 4- to 7-membered heterocyclic group that is         fully saturated, partially unsaturated or fully aromatic and         which heterocyclic group contains one N atom (the atom to which         R^(a) and R^(b) are attached) and, optionally, one or more         further heteroatoms selected from O, S and N, and which         heterocyclic group is optionally substituted by one or more         substituents selected from halo, hydroxy, oxo, C₁₋₄ alkyl, C₁₋₄         alkoxy and C₁₋₄ hydroxyalkyl;         R^(f) represents C₁₋₆ alkyl or C₃₋₇ cycloalkyl;         Het^(x) represents, independently upon each occurrence, a 4- to         7-membered heterocyclic group that is fully saturated, partially         unsaturated or fully aromatic, which group contains one or more         heteroatoms selected from N, O and S, and which group is         optionally substituted by one or more substituents selected from         C₁₋₄ alkyl, halo, hydroxy and oxo;         Het^(y) represents, independently upon each occurrence, a 5- to         10-membered heterocyclic group that is fully aromatic, which         group contains one or more heteroatoms selected from N, O and S,         and which group is optionally substituted by one or more         substituents selected from C₁₋₄ alkyl, halo, hydroxy and oxo;         and         Het^(z) represents, independently upon each occurrence, a 4- to         7-membered heterocyclic group that is fully saturated or         partially unsaturated, which group contains one or more         heteroatoms selected from N, O and S, and which group is         substituted by oxo or hydroxy, and which group is optionally         further substituted by one or more substituents selected from         C₁₋₄ alkyl, halo, hydroxy and oxo,         or a pharmaceutically acceptable salt, solvate or isotopic         derivative thereof,         which compounds may be referred to hereinafter as “the compounds         of the invention”.

Pharmaceutically acceptable salts that may be mentioned include acid addition salts and base addition salts. Such salts may be formed by conventional means, for example by reaction of a free acid or a free base form of a compound of formula I with one or more equivalents of an appropriate acid or base, optionally in a solvent, or in a medium in which the salt is insoluble, followed by removal of said solvent, or said medium, using standard techniques (e.g. in vacuo, by freeze-drying or by filtration). Salts may also be prepared by exchanging a counter-ion of a compound of formula I in the form of a salt with another counter-ion, for example using a suitable ion exchange resin.

Examples of pharmaceutically acceptable salts include acid addition salts derived from mineral acids and organic acids, and salts derived from metals.

For the avoidance of doubt, compounds of formula I may contain the stated atoms in any of their natural or non-natural isotopic forms. In this respect, embodiments of the invention that may be mentioned include those in which:

-   (a) the compound of formula I is not isotopically enriched or     labelled with respect to any atoms of the compound; and -   (b) the compound of formula I is isotopically enriched or labelled     with respect to one or more atoms of the compound.

References herein to an “isotopic derivative” relate to the second of these two embodiments.

In particular embodiments of the invention, the compound of formula I is isotopically enriched or labelled (with respect to one or more atoms of the compound) with one or more stable isotopes. Thus, the compounds of the invention that may be mentioned include, for example, compounds of formula I that are isotopically enriched or labelled with one or more atoms such as deuterium or the like.

Compounds of formula I may exhibit tautomerism. All tautomeric forms and mixtures thereof are included within the scope of the invention.

Unless otherwise specified, alkyl groups and alkoxy groups as defined herein may be straight-chain or, when there is a sufficient number (i.e. a minimum of three) of carbon atoms, be branched. Particular alkyl groups that may be mentioned include, for example, methyl, ethyl, n-propyl, iso-propyl, butyl, n-butyl and tert-butyl. Particular alkoxy groups that may be mentioned include, for example, methoxy, ethoxy, propoxy, and butoxy.

Unless otherwise specified, cycloalkyl groups as defined herein may, when there is a sufficient number (i.e. a minimum of four) of carbon atoms, be part cyclic/acyclic.

Unless otherwise specified, alkylene groups as defined herein may be straight-chain or, when there is a sufficient number (i.e. a minimum of two) of carbon atoms, be branched.

Unless otherwise stated, the point of attachment of aryl groups may be via any atom of the ring system. However, when aryl groups are bicyclic or tricyclic, they are linked to the rest of the molecule via an aromatic ring. C₆₋₁₄ aryl groups include phenyl, naphthyl and the like.

For the avoidance of doubt, oxo substituents that may be present on heterocyclic groups represented by Het^(x), Het^(y), Het^(z), N(R¹)R² and N(R^(a))R^(b) may be attached to any appropriate atoms in the heterocyclic ring including, where valencies allow, to C-, N- and/or S-atoms within the ring (thereby forming keto, N-oxide, S(O) and/or S(O)₂ groups).

Values of Het^(x) that may be mentioned include piperidinyl (e.g. piperidin-4-yl).

Values of Het^(y) that may be mentioned include pyridinyl (e.g. pyridin-2-yl).

Values of Het^(z) that may be mentioned include isothiazolidinyl (e.g. isothiazolidin-2-yl), oxazolidinyl (e.g. oxazolidin-3-yl) or, particularly, piperidinyl (e.g. piperidin-1-yl or piperidin-4-yl), pyrrolidinyl (e.g. pyrrolidin-1-yl) and thiomorpholinyl (e.g. thiomorpholin-4-yl).

Examples of heterocyclic groups that may be formed by R¹, R² and the N-atom to which they are attached include, but are not limited to, azetidin-1-yl, hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl, piperazin-1-yl, piperidin-1-yl, pyrrolidin-1-yl and thiomorpholin-4-yl.

Unless otherwise specified, the term “halo” includes references to fluoro, chloro, bromo or iodo, in particular, to fluoro, chloro or bromo, especially fluoro or chloro.

Embodiments of the invention that may be mentioned include compounds of formula I in which:

R¹ represents R³; R² represents H, R³, C₁₋₆ alkyl or C₃₋₇ cycloalkyl, which latter two groups are optionally substituted by one or more substituents selected from C₁₋₄ alkyl, halo, hydroxy and oxo; or R¹ and R², together with the N-atom to which they are attached, form a 4- to 7-membered heterocyclic group that is fully saturated or partially unsaturated, which heterocyclic group contains one N atom (the atom to which R¹ and R² are attached) and wherein either

-   (a) the heterocyclic group contains one further heteroatom that is S     and is optionally substituted by one or more substituents selected     from halo, hydroxy, oxo, C₁₋₄ alkyl, C₁₋₄ alkoxy and C₁₋₄     hydroxyalkyl or -   (b) the heterocyclic group optionally contains one or more further     heteroatoms selected from N, O and S, is substituted by one or more     substituents selected from C₁₋₄ hydroxyalkyl, C₁₋₄ carboxyalkyl and     —(C₁₋₄ alkylene)-NR^(a)R^(b), and is optionally further substituted     by one or more substituents selected from halo, hydroxy, oxo, CO₂H,     C₁₋₄ alkyl and C₁₋₄ alkoxy; and     R³ represents     -   Het^(x) or     -   C₁₋₆ alkyl substituted by         -   CO₂H,         -   —(OCH₂CH₂)₀₋₄OR^(c),         -   Het^(y) or         -   Het^(z),     -   which C₁₋₆ alkyl group is optionally further substituted by one         or more substituents selected from halo, hydroxy and oxo.

Alternative embodiments of the invention that may be mentioned include those in which one or more of the following definitions apply to the compounds of formula I:

-   (i) R¹ and R², together with the N-atom to which they are attached,     form a 4- to 7-membered heterocyclic group that is fully saturated     or partially unsaturated, which heterocyclic group contains one N     atom (the atom to which R¹ and R² are attached),     -   and wherein the heterocyclic group is fused to a fully saturated         or partially unsaturated 5- or 6-membered heterocyclic ring,         which heterocylic ring contains one or more heteroatoms selected         from from N, O and S and is optionally substituted with one or         more substituents selected from oxo and C₁₋₄ alkyl; -   (ii) R³ represents C₁₋₆ alkyl substituted by     -   NR^(d)C(O)R^(e) or     -   S(O)₁₋₂R^(f),     -   which C₁₋₆ alkyl group is optionally further substituted by one         or more substituents selected from halo, hydroxy and oxo.

Embodiments of the invention that may be mentioned include compounds of formula I in relation to which one or more of the following apply:

-   (i) X represents chloro or, particularly, fluoro; -   (ii) Ak represents C₁₋₄ alkyl optionally substituted by one or more     fluorine atoms; -   (iii) s represents 1; -   (iv) L represents C(O); -   (v) Q represents a 6-membered heteroaromatic ring containing 2 N     atoms, such as pyrazinyl; -   (vi) R¹ represents R³; -   (vii) R² represents H, C₁₋₃ alkyl or R³; -   (viii) R¹ and R², together with the N-atom to which they are     attached, form a 4- to 6-membered heterocyclic group that is fully     saturated, which heterocyclic group contains one N atom (the atom to     which R¹ and R² are attached) and wherein     -   (a) the heterocyclic group contains one further heteroatom that         is S and is optionally substituted by one or more substituents         selected from halo, hydroxy, oxo, C₁₋₂ alkyl, C₁₋₂ alkoxy and         C₁₋₂ hydroxyalkyl, or     -   (b) the heterocyclic group optionally contains one further         heteroatom selected from N, O and S, is substituted by a         substituent selected from C₁₋₃ hydroxyalkyl, C₁₋₃ carboxyalkyl         and —(C₁₋₃ alkylene)-NR^(a)R^(b), and is optionally further         substituted by one or more hydroxy or C₁₋₂ alkyl groups, or     -   (c) the heterocyclic group is fused to a fully saturated 5- or         6-membered heterocyclic ring, which heterocylic ring contains         one or two heteroatoms selected from from N, O and S and is         optionally substituted by one to three substituents selected         from oxo and C₁₋₂ alkyl     -   (e.g. R¹ and R², together with the N-atom to which they are         attached, form a 5- or 6-membered heterocyclic group that is         fully saturated, which heterocyclic group contains one N atom         (the atom to which R¹ and R² are attached) and wherein either     -   (a) the heterocyclic group contains one further heteroatom that         is S and is optionally substituted by one or more substituents         selected from halo, hydroxy, oxo, C₁₋₂ alkyl, C₁₋₂ alkoxy and         C₁₋₂ hydroxyalkyl or     -   (b) the heterocyclic group optionally contains one further         heteroatom selected from N, O and S, is substituted by a         substituent selected from C₁₋₃ hydroxyalkyl, C₁₋₃ carboxyalkyl         and —(C₁₋₃ alkylene)-NR^(a)R^(b), and is optionally further         substituted by C₁₋₂ alkyl); -   (ix) R³ represents     -   Het^(x) or     -   C₁₋₄ alkyl substituted by (e.g. C₂-3 n-alkyl terminated by)         -   CO₂H,         -   —(OCH₂CH₂)₀₋₃OR^(c),         -   Het^(y),         -   Het^(z),         -   NR^(d)C(O)R^(e) or         -   S(O)₁₋₂R^(f), -    (e.g. R³ represents     -   Het^(x) or     -   C₁₋₄ alkyl substituted by         -   CO₂H,         -   —(OCH₂CH₂)₀₋₁OR^(c),         -   Het^(y) or         -   Het^(z)); -   (x) R^(a) and R^(b) represent H or C₁₋₂ alkyl, or R^(a) and R^(b),     together with the N-atom to which they are attached, form a 5- or     6-membered heterocyclic group that is fully saturated, partially     unsaturated or fully aromatic and which heterocyclic group contains     one N atom (the atom to which R^(a) and R^(b) are attached) and,     optionally, one or two further heteroatoms selected from O, S and N,     and which heterocyclic group is optionally substituted by one or     more substituents selected from hydroxy, oxo, C₁₋₂ alkyl, C₁₋₂     alkoxy and C₁₋₂ hydroxyalkyl; -   (xi) R^(c) represents H or C₁₋₂ alkyl; -   (xii) R^(d) and R^(e) independently represent H or, particularly,     C₁₋₂ alkyl; -   (xiii) R^(f) represents C₁₋₂ alkyl; -   (xiv) Het^(x) represents, independently upon each occurrence, a 5-     or 6-membered heterocyclic group that is fully saturated, which     group contains one or two heteroatoms selected from N, O and S, and     which group is optionally substituted by one to three substituents     selected from C₁₋₂ alkyl, halo, hydroxy and oxo; -   (xv) Het^(y) represents, independently upon each occurrence, a 5- or     6-membered heterocyclic group that is fully aromatic, which group     contains one to three heteroatoms selected from N, O and S and which     group is optionally substituted by one or more substituents selected     from C₁₋₂ alkyl, halo, hydroxy and oxo; -   (xvi) Het^(z) represents, independently upon each occurrence, a 5-     or 6-membered heterocyclic group that is fully saturated, which     group contains one or two heteroatoms selected from N, O and S, and     which group is substituted by oxo or hydroxy, and which group is     optionally further substituted by one or more substituents selected     from C₁₋₂ alkyl, halo, hydroxy and oxo.

Embodiments of the invention that may be mentioned include those in which the compound of formula I is a compound of formula Ia or Ib,

wherein X, Ak, L, Q, R¹ and R² are as hereinbefore defined.

Embodiments of the invention that may be mentioned include those in which one or more of the following definitions apply to the compounds of formula I, la or Ib:

-   (i) X represents fluoro; -   (ii) Ak represents C₁₋₃ alkyl, such as ethyl; -   (iii) L represents C(O); -   (iv) Q represents pyrazinyl; -   (v) R¹ represents R³; -   (vi) R² represents H, methyl or R³; -   (vii) R¹ and R², together with the N-atom to which they are     attached, form a 5- or 6-membered heterocyclic group that is fully     saturated, which heterocyclic group contains one N atom (the atom to     which R¹ and R² are attached) and wherein either     -   (a) the heterocyclic group contains one further heteroatom that         is S and is optionally substituted by one or more substituents         selected from hydroxy, oxo and methyl or     -   (b) the heterocyclic group optionally contains one further         heteroatom selected from N, O and S, is substituted by a         substituent selected from C₁₋₂ hydroxyalkyl (e.g. hydroxymethyl)         and —(C₁₋₃ alkylene)-NR^(a)R^(b), and is optionally further         substituted by methyl; -   (viii) R³ represents     -   Het^(x) or     -   C₁₋₄ alkyl substituted by         -   CO₂H (e.g. to form —(CH₂)₃—CO₂H),         -   —(OCH₂CH₂)₀₋₂OR^(c) (e.g. to form —(CH₂)₂—(OCH₂CH₂)₂OR^(c)             or —(CH₂)₂—OR^(c)),         -   Het^(y) (e.g. to form —(CH₂)₁₋₂-Het^(y)),         -   Het^(z) (e.g. to form —(CH₂)₁₋₃-Het^(z)) or         -   NR^(d)C(O)R^(e) (e.g. to form —(CH₂)₂—NR^(d)C(O)R^(e)) -    (e.g. R³ represents     -   Het^(x) or     -   C₁₋₄ alkyl substituted by         -   CO₂H (e.g. to form —(CH₂)₃—CO₂H),         -   —OR^(c) (e.g. to form —(CH₂)₂—OR^(c)),         -   Het^(y) (e.g. to form —(CH₂)₁₋₂-Het^(y)) or         -   Het^(z) (e.g. to form —(CH₂)₁₋₃-Het^(z))); -   (ix) R^(a) and R^(b) represent H or methyl, or R^(a) and R^(b),     together with the N-atom to which they are attached, form a 5- or     6-membered heterocyclic group that is fully aromatic and which     heterocyclic group contains one N atom (the atom to which R^(a) and     R^(b) are attached) and, optionally, one or two further heteroatoms     selected from O, S and N, and which heterocyclic group is optionally     substituted by one or more substituents selected from methyl,     methoxy and hydroxymethyl; -   (x) R^(c) represents H or methyl; -   (xi) R^(d) and R^(e) independently represent H or, particularly,     methyl; -   (xii) Het^(x) represents, independently upon each occurrence, a 5-     or 6-membered heterocyclic group that is fully saturated, which     group contains one or two heteroatoms selected from N, O and S, and     which group is optionally substituted by one or two substituents     selected from methyl, hydroxy and oxo; -   (xiii) Het^(y) represents, independently upon each occurrence, a 5-     or 6-membered heterocyclic group that is fully aromatic, which group     contains one or two heteroatoms selected from N, O and S and which     group is optionally substituted by one or two methyl substituents; -   (xiv) Het^(z) represents, independently upon each occurrence, a 5-     or 6-membered heterocyclic group that is fully saturated, which     group contains one or two heteroatoms selected from N, O and S, and     which group is substituted by oxo or hydroxy, and which group is     optionally further substituted by one or more substituents selected     from methyl, hydroxy and oxo.

Further embodiments of the invention that may be mentioned include those in which the compound of formula I is a compound of formula Ic,

wherein X, Ak, R¹ and R² are as hereinbefore defined.

Embodiments of the invention that may be mentioned include those in which one or more of the following definitions apply to the compounds of formula I, Ia, Ib or Ic:

-   (i) X represents fluoro; -   (ii) Ak represents ethyl; -   (iii) R¹ represents R³; -   (iv) R² represents H, methyl or R³ (e.g. R² represents methyl or     R³); -   (v) R¹ and R², together with the N-atom to which they are attached,     form a 5- or 6-membered heterocyclic group that is fully saturated,     which heterocyclic group contains one N atom (the atom to which R¹     and R² are attached) and wherein either     -   (a) the heterocyclic group contains one further heteroatom that         is S and the heterocyclic group is optionally substituted by oxo         or     -   (b) the heterocyclic group is substituted by C₁₋₂ hydroxyalkyl         (e.g. hydroxymethyl); -   (vi) R³ represents C₁₋₃ alkyl (e.g. ethyl) substituted by     -   —(OCH₂CH₂)₀₋₂OR^(c) (e.g. to form —(CH₂)₂—(OCH₂CH₂)₂OR^(c) or         —(CH₂)₂—OR^(c)),     -   Het^(z) (e.g. to form —(CH₂)₂₋₃-Het^(z)) or     -   NCH₃C(O)CH₃ (e.g. to form —(CH₂)₂—NCH₃C(O)CH₃ -    (e.g. R³ represents C₁₋₃ alkyl (e.g. ethyl) substituted by     -   —OR^(c) (e.g. to form —(CH₂)₂—OR^(c)) or     -   Het^(z) (e.g. to form —(CH₂)₂₋₃-Het^(z))); -   (vii) R^(c) represents methyl; -   (viii) Het^(z) represents a 5- or 6-membered heterocyclic group that     is fully saturated, which group contains one or two heteroatoms     selected from N, O and S, and which group is substituted by oxo and     is optionally further substituted by a substituent selected from     methyl and oxo (e.g. Het^(z) represents pyrrolidinyl or     isothiazolidinyl, such as pyrrolidin-1-yl substituted by oxo (e.g.     in the 2-position) or isothiazolidin-2-yl disubstituted by oxo in     the 1-position) -    (e.g. Het^(z) represents a 5- or 6-membered heterocyclic group that     is fully saturated, which group contains one or two heteroatoms     (e.g. one heteroatom) selected from N, O and S, and which group is     substituted by oxo (e.g. Het^(z) represents pyrrolidinyl, such as     pyrrolidin-1-yl, substituted by oxo (e.g. in the 2-position))).

Other compounds of formula I, Ia, Ib and Ic that may be mentioned include the compounds of the examples described hereinafter. Thus, embodiments of the invention that may be mentioned include those in which the compound of formula I, Ia, Ib and Ic is a compound selected from the list:

-   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(methyl(1-methylpiperidin-4-yl)amino)pyrazin-2-yl)methanone; -   4-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)butanoic     acid; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(4-hydroxypiperidin-1-yl)ethyl)amino)pyrazin-2-yl)methanone; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(methyl(pyridin-2-ylmethyl)amino)     pyrazin-2-yl)methanone; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(1-oxidothiomorpholino)ethyl)amino)     pyrazin-2-yl)methanone; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(1-oxidothiomorpholino)     pyrazin-2-yl)methanone; -   (5-(4-(2-(dimethylamino)ethyl)piperazin-1-yl)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(4-hydroxy-1-methylpiperidin-4-yl)ethyl)amino)pyrazin-2-yl)methanone; -   (S)-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(2-(hydroxymethyl)pyrrolidin-1-yl)pyrazin-2-yl)methanone; -   (5-(bis(2-methoxyethyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone; -   (5-(4-(2-(1H-imidazol-1-yl)ethyl)piperazin-1-yl)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone; -   1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)     pyrazin-2-yl)(methyl)amino)ethyl)pyrrolidin-2-one; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-(4-(2-hydroxyethyl)piperazin-1-yl)pyrazin-2-yl)methanone; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino)     pyrazin-2-yl)methanone; -   N-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)ethyl)-N-methylacetamide; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-(3-(hydroxymethyl)azetidin-1-yl)pyrazin-2-yl)methanone; -   (5-(bis(2-hydroxyethyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)methanone; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2-(methylsulfonyl)ethyl)amino)     pyrazin-2-yl)methanone; -   1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)amino)ethyl)pyrrolidin-2-one; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidin-1-yl)pyrazin-2-yl)methanone; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)pyrazin-2-yl)methanone; -   3-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)     pyrazin-2-yl)(methyl)amino)ethyl)oxazolidin-2-one; -   (5-((2-(1,1-dioxidoisothiazolidin-2-yl)ethyl)(methyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)methanone;     and -   (3aR,6aS)-5-(5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)     pyrazin-2-yl)tetrahydropyrrolo[3,4-c]pyrrole-1,3(2H,3aH)-dione,     or a pharmaceutically acceptable salt, solvate or isotopic     derivative thereof.

Further embodiments of the invention that may be mentioned include those in which either

-   (i) the compound of formula I, Ia, Ib or Ic represents, or -   (ii) the compound of formula I, Ia, Ib or Ic is as hereinbefore     defined, provided that is does not represent -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(1-oxidothiomorpholino)     pyrazin-2-yl)methanone; -   1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)     pyrazin-2-yl)(methyl)amino)ethyl)pyrrolidin-2-one; -   (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino)     pyrazin-2-yl)methanone; -   N-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)     pyrazin-2-yl)(methyl)amino)ethyl)-N-methylacetamide; and/or -   (5-((2-(1,1-dioxidoisothiazolidin-2-yl)ethyl)(methyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)methanone;     or a pharmaceutically acceptable salt, solvate or isotopic     derivative thereof -   (e.g.     (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(1-oxidothiomorpholino)pyrazin-2-yl)methanone;     and/or -   1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)     pyrazin-2-yl)(methyl)amino)ethyl)pyrrolidin-2-one, or a     pharmaceutically acceptable salt, solvate or isotopic derivative     thereof).

Examples of salts of compounds of formula I, Ia, Ib and Ic include all pharmaceutically acceptable salts, such as, without limitation, acid addition salts of strong mineral acids such as HCl, H₂SO₄ and HBr salts (e.g. HCl or HBr salts) and addition salts of strong organic acids such as methanesulfonic acid.

References herein to a compound of the invention (a compound of formula I, Ia, Ib or Ic) are intended to include references to the compound and to all pharmaceutically acceptable salts, solvates and/or tautomers of said compound, unless the context specifically indicates otherwise. In this respect, solvates that may be mentioned include hydrates.

The compounds of the invention (compounds of formula I, Ia, Ib and Ic) are JAK kinase inhibitors and are therefore useful in medicine, in particular for the treatment of inflammatory diseases. Further aspects of the invention that may be mentioned therefore include the following.

-   (a) A pharmaceutical formulation comprising a compound of formula I,     Ia, Ib or Ic, as hereinbefore defined, or pharmaceutically     acceptable salt, solvate or isotopic derivative thereof, in     admixture with a pharmaceutically acceptable adjuvant, diluent or     carrier. -   (b) A combination product comprising     -   (A) a compound of formula I, Ia, Ib or Ic, as hereinbefore         defined, or pharmaceutically acceptable salt, solvate or         isotopic derivative thereof, and     -   (B) another therapeutic agent, wherein each of components (A)         and (B) is formulated in admixture with a         pharmaceutically-acceptable adjuvant, diluent or carrier.     -   In this aspect of the invention, the combination product may be         either a single (combination) pharmaceutical formulation or a         kit-of-parts.     -   Thus, this aspect of the invention encompasses a pharmaceutical         formulation including a compound of formula I, Ia, Ib or Ic, as         hereinbefore defined, or pharmaceutically acceptable salt,         solvate or isotopic derivative thereof, and another therapeutic         agent, in admixture with a pharmaceutically acceptable adjuvant,         diluent or carrier (which formulation is hereinafter referred to         as a “combined preparation”).

It also encompasses a kit of parts comprising components:

-   -   (i) a pharmaceutical formulation including a compound of formula         I, Ia, Ib or Ic, as hereinbefore defined, or pharmaceutically         acceptable salt, solvate or isotopic derivative thereof, in         admixture with a pharmaceutically acceptable adjuvant, diluent         or carrier; and     -   (ii) a pharmaceutical formulation including another therapeutic         agent, in admixture with a pharmaceutically-acceptable adjuvant,         diluent or carrier,     -   which components (i) and (ii) are each provided in a form that         is suitable for administration in conjunction with the other.     -   Component (i) of the kit of parts is thus component (A) above in         admixture with a pharmaceutically acceptable adjuvant, diluent         or carrier. Similarly, component (ii) is component (B) above in         admixture with a pharmaceutically acceptable adjuvant, diluent         or carrier.

-   (c) A process for preparing the pharmaceutical formulation of     aspect (a) above, said process comprising the step of admixing the     compound of formula I, Ia, Ib or Ic, as hereinbefore defined, or     pharmaceutically acceptable salt, solvate or isotopic derivative     thereof, with a pharmaceutically acceptable adjuvant, diluent or     carrier.     -   Embodiments of this aspect of the invention that may be         mentioned include those in which the pharmaceutically acceptable         adjuvant, diluent or carrier is a topically acceptable adjuvant,         diluent or carrier (and/or wherein the process is for preparing         a topical pharmaceutical formulation, i.e. a pharmaceutical         formulation that is adapted for topical administration).

-   (d) A compound of formula I, Ia, Ib or Ic, as hereinbefore defined,     or pharmaceutically acceptable salt, solvate or isotopic derivative     thereof, for use in medicine (or for use as a medicament or as a     pharmaceutical).

-   (e) A compound of formula I, Ia, Ib or Ic, as hereinbefore defined,     or pharmaceutically acceptable salt, solvate or isotopic derivative     thereof, or a pharmaceutical formulation or combination product as     defined in connection with aspect (a) or (b) of the invention, for     use in the treatment or prevention of an inflammatory disease.

-   (f) The use of     -   a compound of formula I, Ia, Ib or Ic, as hereinbefore defined,         or pharmaceutically acceptable salt, solvate or isotopic         derivative thereof, or     -   a pharmaceutical formulation or combination product as defined         in connection with aspect (a) or (b) of the invention,

-    for the preparation of a medicament for the treatment or prevention     of an inflammatory disease.

-   (g) A method of treating or preventing an inflammatory disease, said     method comprising administering to a subject an effective amount of     -   a compound of formula I, Ia, Ib or Ic, as hereinbefore defined,         or pharmaceutically acceptable salt, solvate or isotopic         derivative thereof, or     -   a pharmaceutical formulation or combination product as defined         in connection with aspect (a) or (b) of the invention.

-   (h) A method of sensitizing a subject to the anti-inflammatory     effects of a corticosteroid, said method comprising administering to     the subject an effective amount of     -   a compound of formula I, Ia, Ib or Ic, as hereinbefore defined,         or pharmaceutically acceptable salt, solvate or isotopic         derivative thereof, or     -   a pharmaceutical formulation or combination product as defined         in connection with aspect (a) or (b) of the invention.     -   Embodiments of this aspect of the invention that may be         mentioned include those in which the subject is one who has         become refractory to the anti-inflammatory effects of a         corticosteroid.

References herein to “preventing an inflammatory disease” include references to preventing (or reducing the likelihood of) the recurrence of an inflammatory disease in a subject who has previously suffered from such a disease (e.g. a subject who has previously received treatment for that disease, for example treatment according to the method described in (g) above).

Thus, still further aspects of the invention that may be mentioned include the following.

-   (i) A compound of formula I, Ia, Ib or Ic, as hereinbefore defined,     or pharmaceutically acceptable salt, solvate or isotopic derivative     thereof, or a pharmaceutical formulation or combination product as     defined in connection with aspect (a) or (b) of the invention, for     use in reducing the likelihood of the recurrence of an inflammatory     disease in a subject who has previously received treatment for that     disease (e.g. treatment with a compound of formula I, Ia, Ib or Ic,     as hereinbefore defined, or pharmaceutically acceptable salt,     solvate or isotopic derivative thereof, or a pharmaceutical     formulation or combination product as defined in connection with     aspect (a) or (b) of the invention). -   (j) The use of     -   a compound of formula I, Ia, Ib or Ic, as hereinbefore defined,         or pharmaceutically acceptable salt, solvate or isotopic         derivative thereof, or     -   a pharmaceutical formulation or combination product as defined         in connection with aspect (a) or (b) of the invention, -    for the preparation of a medicament for reducing the likelihood of     the recurrence of an inflammatory disease in a subject who has     previously received treatment for that disease (e.g. treatment with     a compound of formula I, Ia, Ib or Ic, as hereinbefore defined, or     pharmaceutically acceptable salt, solvate or isotopic derivative     thereof, or a pharmaceutical formulation or combination product as     defined in connection with aspect (a) or (b) of the invention). -   (k) A method of reducing the likelihood of the recurrence of an     inflammatory disease in a subject who has previously received     treatment for that disease (e.g. treatment with a compound of     formula I, Ia, Ib or Ic, as hereinbefore defined, or     pharmaceutically acceptable salt, solvate or isotopic derivative     thereof, or a pharmaceutical formulation or combination product as     defined in connection with aspect (a) or (b) of the invention), said     method comprising administering to said subject an effective amount     of     -   a compound of formula I, Ia, Ib or Ic, as hereinbefore defined,         or pharmaceutically acceptable salt, solvate or isotopic         derivative thereof, or     -   a pharmaceutical formulation or combination product as defined         in connection with aspect (a) or (b) of the invention.

Formulations

In relation to aspects (a) and (b) above, diluents and carriers that may be mentioned include those suitable for parenteral, oral, topical, mucosal and rectal administration.

The pharmaceutical formulations and combination products of aspects (a) and (b) above may be prepared e.g. for parenteral, subcutaneous, intramuscular, intravenous, intra-articular, intravitreous, periocular, retrobulbar, subconjunctival, sub-Tenon, topical ocular or peri-articular administration, particularly in the form of liquid solutions, emulsions or suspensions; for oral administration, particularly in the form of tablets or capsules, and especially involving technologies aimed at furnishing colon-targeted drug release (Patel, M. M. Expert Opin. Drug Deliv. 2011, 8 (10), 1247-1258); for topical e.g. pulmonary or intranasal administration, particularly in the form of powders, nasal drops or aerosols and transdermal administration; for topical ocular administration, particularly in the form of solutions, emulsions, suspensions, ointments, implants/inserts, gels, jellies or liposomal microparticle formulations (Ghate, D.; Edelhauser, H. F. Expert Opin. Drug Deliv. 2006, 3 (2), 275-287); for ocular administration, particularly in the form of biodegradable and non-biodegradable implants, liposomes and nanoparticles (Thrimawithana, T. R. et al. Drug Discov. Today 2011, 16 (5/6), 270-277); for mucosal administration e.g. to buccal, sublingual or vaginal mucosa, and for rectal administration e.g. in the form of a suppository or enema.

The pharmaceutical formulations and combination products of aspects (a) and (b) above may conveniently be administered in unit dosage form and may be prepared by any of the methods well-known in the pharmaceutical art, for example as described in Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Company, Easton, Pa., (1985). Formulations for parenteral administration may contain as excipients sterile water or saline, alkylene glycols such as propylene glycol, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, hydrogenated naphthalenes and the like. Formulations for nasal administration may be solid and may contain excipients, for example, lactose or dextran, or may be aqueous or oily solutions for use in the form of nasal drops or metered sprays. For buccal administration, typical excipients include sugars, calcium stearate, magnesium stearate, pregelatinised starch, and the like.

Pharmaceutical formulations and combination products suitable for oral administration may comprise one or more physiologically compatible carriers and/or excipients and may be in solid or liquid form. Tablets and capsules may be prepared with binding agents, for example, syrup, acacia, gelatin, sorbitol, tragacanth, or poly-vinylpyrrolidone; fillers, such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, or glycine; lubricants, such as magnesium stearate, talc, polyethylene glycol, or silica; and surfactants, such as sodium lauryl sulfate. Liquid compositions may contain conventional additives such as suspending agents, for example sorbitol syrup, methyl cellulose, sugar syrup, gelatin, carboxymethyl-cellulose, or edible fats; emulsifying agents such as lecithin, or acacia; vegetable oils such as almond oil, coconut oil, cod liver oil, or peanut oil; preservatives such as butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT). Liquid compositions may be encapsulated in, for example, gelatin to provide a unit dosage form.

Solid oral dosage forms include tablets, two-piece hard shell capsules and soft elastic gelatin (SEG) capsules. Such two-piece hard shell capsules may be made from, for example, gelatin or hydroxylpropyl methylcellulose (HPMC).

A dry shell formulation typically comprises of about 40% to 60% w/w concentration of gelatin, about a 20% to 30% concentration of plasticizer (such as glycerin, sorbitol or propylene glycol) and about a 30% to 40% concentration of water. Other materials such as preservatives, dyes, opacifiers and flavours also may be present. The liquid fill material comprises a solid drug that has been dissolved, solubilized or dispersed (with suspending agents such as beeswax, hydrogenated castor oil or polyethylene glycol 4000) or a liquid drug in vehicles or combinations of vehicles such as mineral oil, vegetable oils, triglycerides, glycols, polyols and surface-active agents.

A compound of the invention may be administered topically (e.g. to the lung, eye or intestines). Thus, embodiments of aspects (a) and (b) above that may be mentioned include pharmaceutical formulations and combination products that are adapted for topical administration. Such formulations include those in which the excipients (including any adjuvant, diluent and/or carrier) are topically acceptable.

Topical administration to the lung may be achieved by use of an aerosol formulation. Aerosol formulations typically comprise the active ingredient suspended or dissolved in a suitable aerosol propellant, such as a chlorofluorocarbon (CFC) or a hydrofluorocarbon (HFC). Suitable CFC propellants include trichloromonofluoromethane (propellant 11), dichlorotetrafluoroethane (propellant 114), and dichlorodifluoromethane (propellant 12). Suitable HFC propellants include tetrafluoroethane (HFC-134a) and heptafluoropropane (HFC-227). The propellant typically comprises 40% to 99.5% e.g. 40% to 90% by weight of the total inhalation composition. The formulation may comprise excipients including co-solvents (e.g. ethanol) and surfactants (e.g. lecithin, sorbitan trioleate and the like). Other possible excipients include polyethylene glycol, polyvinylpyrrolidone, glycerine and the like. Aerosol formulations are packaged in canisters and a suitable dose is delivered by means of a metering valve (e.g. as supplied by Bespak, Valois or 3M or alternatively by Aptar, Coster or Vari).

Topical administration to the lung may also be achieved by use of a non-pressurised formulation such as an aqueous solution or suspension. This may be administered by means of a nebuliser e.g. one that can be hand-held and portable or for home or hospital use (i.e. non-portable). The formulation may comprise excipients such as water, buffers, tonicity adjusting agents, pH adjusting agents, surfactants and co-solvents. Suspension liquid and aerosol formulations (whether pressurised or unpressurised) will typically contain the compound of the invention in finely divided form, for example with a D₅₀ of 0.5-10 μm e.g. around 1-5 μm. Particle size distributions may be represented using D₁₀, D₅₀ and D₉₀ values. The D₅₀ median value of particle size distributions is defined as the particle size in microns that divides the distribution in half. The measurement derived from laser diffraction is more accurately described as a volume distribution, and consequently the D₅₀ value obtained using this procedure is more meaningfully referred to as a Dv₅₀ value (median for a volume distribution). As used herein Dv values refer to particle size distributions measured using laser diffraction. Similarly, D₁₀ and D₉₀ values, used in the context of laser diffraction, are taken to mean Dv₁₀ and Dv₉₀ values and refer to the particle size whereby 10% of the distribution lies below the D₁₀ value, and 90% of the distribution lies below the D₉₀ value, respectively.

Topical administration to the lung may also be achieved by use of a dry-powder formulation. A dry powder formulation will contain the compound of the disclosure in finely divided form, typically with a mass mean aerodynamic diameter (MMAD) of 1-10 μm or a D₅₀ of 0.5-10 μm e.g. around 1-5 μm. Powders of the compound of the invention in finely divided form may be prepared by a micronization process or similar size reduction process. Micronization may be performed using a jet mill such as those manufactured by Hosokawa Alpine. The resultant particle size distribution may be measured using laser diffraction (e.g. with a Malvern Mastersizer 2000S instrument). The formulation will typically contain a topically acceptable diluent such as lactose, glucose or mannitol (preferably lactose), usually of large particle size e.g. an MMAD of 50 μm or more, e.g. 100 μm or more or a D₅₀ of 40-150 μm. As used herein, the term “lactose” refers to a lactose-containing component, including α-lactose monohydrate, β-lactose monohydrate, α-lactose anhydrous, β-lactose anhydrous and amorphous lactose. Lactose components may be processed by micronization, sieving, milling, compression, agglomeration or spray drying. Commercially available forms of lactose in various forms are also encompassed, for example Lactohale® (inhalation grade lactose; DFE Pharma), InhaLac®70 (sieved lactose for dry powder inhaler; Meggle), Pharmatose® (DFE Pharma) and Respitose® (sieved inhalation grade lactose; DFE Pharma) products. In one embodiment, the lactose component is selected from the group consisting of α-lactose monohydrate, α-lactose anhydrous and amorphous lactose. Preferably, the lactose is α-lactose monohydrate.

Dry powder formulations may also contain other excipients such as sodium stearate, calcium stearate or magnesium stearate.

A dry powder formulation is typically delivered using a dry powder inhaler (DPI) device. Examples of dry powder delivery systems include SPINHALER, DISKHALER, TURBOHALER, DISKUS and CLICKHALER. Further examples of dry powder delivery systems include ECLIPSE, NEXT, ROTAHALER, HANDIHALER, AEROLISER, CYCLOHALER, BREEZHALER/NEOHALER, MONODOSE, FLOWCAPS, TWINCAPS, X-CAPS, TURBOSPIN, ELPENHALER, MIATHALER, TWISTHALER, NOVOLIZER, PRESSAIR, ELLIPTA, ORIEL dry powder inhaler, MICRODOSE, PULVINAL, EASYHALER, ULTRAHALER, TAIFUN, PULMOJET, OMNIHALER, GYROHALER, TAPER, CONIX, XCELOVAIR and PROHALER.

In one embodiment a compound of the present invention is provided in a micronized dry powder formulation, for example further comprising lactose of a suitable grade optionally together with magnesium stearate, filled into a single dose device such as AEROLISER or filled into a multi dose device such as DISKUS.

The compounds of the present invention may also be administered rectally, for example in the form of suppositories or enemas, which include aqueous or oily solutions as well as suspensions and emulsions. Such compositions are prepared following standard procedures, well known by those skilled in the art. For example, suppositories can be prepared by mixing the active ingredient with a conventional suppository base such as cocoa butter or other glycerides, e.g. Suppocire. In this case, the drug is mixed with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

Generally, for compositions intended to be administered topically to the eye in the form of eye drops or eye ointments, the total amount of the inhibitor will be about 0.0001 to less than 4.0% (w/w).

Preferably, for topical ocular administration, the compositions administered according to the present invention will be formulated as solutions, suspensions, emulsions and other dosage forms. Aqueous solutions are generally preferred, based on ease of formulation, as well as a patient's ability to administer such compositions easily by means of instilling one to two drops of the solutions in the affected eyes. However, the compositions may also be suspensions, viscous or semi-viscous gels, or other types of solid or semi-solid compositions. Suspensions may be preferred for compounds that are sparingly soluble in water.

The compositions administered according to the present invention may also include various other ingredients, including, but not limited to, tonicity agents, buffers, surfactants, stabilizing polymer, preservatives, co-solvents and viscosity building agents. Preferred pharmaceutical compositions of the present invention include the inhibitor with a tonicity agent and a buffer. The pharmaceutical compositions of the present invention may further optionally include a surfactant and/or a palliative agent and/or a stabilizing polymer.

Various tonicity agents may be employed to adjust the tonicity of the composition, preferably to that of natural tears for ophthalmic compositions. For example, sodium chloride, potassium chloride, magnesium chloride, calcium chloride, simple sugars, such as dextrose, fructose, galactose, and/or simply polyols, such as the sugar alcohols mannitol, sorbitol, xylitol, lactitol, isomaltitol, maltitol, and hydrogenated starch hydrolysates may be added to the composition to approximate physiological tonicity. Such an amount of tonicity agent will vary, depending on the particular agent to be added. In general, however, the compositions will have a tonicity agent in an amount sufficient to cause the final composition to have an ophthalmically acceptable osmolality (generally about 150-450 mOsm, preferably 250-350 mOsm and most preferably at approximately 290 mOsm). In general, the tonicity agents of the invention will present in the range of 2 to 5% w/w. Preferred tonicity agents of the invention include the simple sugars or the sugar alcohols, such as D-mannitol.

An appropriate buffer system (e.g. sodium phosphate, sodium acetate, sodium citrate, sodium borate or boric acid) may be added to the compositions to prevent pH drift under storage conditions. The particular concentration will vary, depending on the agent employed. Preferably however, the buffer will be chosen to maintain a target pH within the range of pH 5 to 8.

Surfactants may optionally be employed to deliver higher concentrations of inhibitor. The surfactants function to solubilise the inhibitor and stabilise colloid dispersion, such as micellar solution, microemulsion, emulsion and suspension. Examples of surfactants which may optionally be used include polysorbate, poloxamer, polyoxyl 40 stearate, polyoxyl castor oil, tyloxapol, triton, and sorbitan monolaurate. Preferred surfactants to be employed in the invention have a hydrophile/lipophile/balance “HLB” in the range of 12.4 to 13.2 and are acceptable for ophthalmic use, such as TritonX114 and tyloxapol.

Additional agents that may be added to the ophthalmic compositions of the present invention are demulcents which function as a stabilising polymer. The stabilizing polymer should be an ionic/charged example with precedence for topical ocular use, more specifically, a polymer that carries negative charge on its surface that can exhibit a zeta-potential of (−)10-50 mV for physical stability and capable of making a dispersion in water (i.e. water soluble). A preferred stabilising polymer of the invention would be polyelectrolyte, or polyelectrolytes if more than one, from the family of cross-linked polyacrylates, such as carbomers, polycarbophil and Pemulen®, specifically Carbomer 974p (polyacrylic acid), at 0.1-0.5% w/w.

Other compounds may also be added to the ophthalmic compositions of the present invention to increase the viscosity of the carrier. Examples of viscosity enhancing agents include, but are not limited to: polysaccharides, such as hyaluronic acid and its salts, chondroitin sulfate and its salts, dextrans, various polymers of the cellulose family, vinyl polymers and acrylic acid polymers.

Topical ophthalmic products are typically packaged in multidose form. Preservatives are thus required to prevent microbial contamination during use. Suitable preservatives include: benzalkonium chloride, chlorobutanol, benzododecinium bromide, methyl paraben, propyl paraben, phenylethyl alcohol, edentate disodium, sorbic acid, polyquaternium-1, or other agents known to those skilled in the art. Such preservatives are typically employed at a level of from 0.001 to 1.0% w/v. Unit dose compositions of the present invention will be sterile, but typically unpreserved. Such compositions, therefore, generally will not contain preservatives.

The medical practitioner, or other skilled person, will be able to determine a suitable dosage for the compounds of the invention, and hence the amount of the compound of the invention that should be included in any particular pharmaceutical formulation (whether in unit dosage form or otherwise).

Embodiments of the invention that may be mentioned in connection with the combination products described at (b) above include those in which the other therapeutic agent is one or more therapeutic agents that are known by those skilled in the art to be suitable for treating inflammatory diseases (e.g. the specific diseases mentioned below).

For example, for the treatment of respiratory disorders (such as COPD or asthma), the other therapeutic agent is one or more agents selected from the list comprising:

-   -   steroids (e.g. budesonide, beclomethasone dipropionate,         fluticasone propionate, mometasone furoate, fluticasone furoate;         a further example is ciclesonide);     -   beta agonists, particularly beta2 agonists (e.g. terbutaline,         salbutamol, salmeterol, formoterol; further examples are         vilanterol, olodaterol, reproterol and fenoterol); and     -   xanthines (e.g. theophylline).

For example, for the treatment of respiratory disorders (such as COPD or asthma), the other therapeutic agent is one or more agents selected from the list comprising:

-   -   muscarinic antagonists (e.g. tiotropium, umeclidinium,         glycopyrronium, aclidinium and daratropium, any of these for         example as the bromide salt); and     -   phosphodiesterase inhibitors.

Further, for the treatment of gastrointestinal disorders (such as Crohn's disease or ulcerative colitis), the other therapeutic agent may be, for example, one or more agents selected from the list comprising:

-   -   5-aminosalicylic acid, or a prodrug thereof (such as         sulfasalazine, olsalazine or balsalazide);     -   corticosteroids (e.g. prednisolone, methylprednisolone, or         budesonide);     -   immunosuppressants (e.g. cyclosporin, tacrolimus, methotrexate,         azathioprine or 6-mercaptopurine);     -   anti-TNFα antibodies (e.g. infliximab, adalimumab, certolizumab         pegol or golimumab); anti-IL12/IL23 antibodies (e.g.         ustekinumab) or small molecule IL12/IL23 inhibitors (e.g.         apilimod);     -   anti-αβ47 antibodies (e.g. vedolizumab);     -   toll-like receptor (TLR) blockers (e.g. BL-7040; Avecia         (Cambridge, UK)); MAdCAM-1 blockers (e.g. PF-00547659);     -   antibodies against the cell adhesion molecule α4-integrin (e.g.         natalizumab);     -   antibodies against the IL2 receptor a subunit (e.g. daclizumab         or basiliximab);     -   anti-Smad7 antibodies (e.g. mongersen (GED0301;         all-P-ambo-2′-deoxy-P-thioguanylyl-(3′→5′)-P-thiothymidylyl-(3′→5′)-2′-deoxy-5-methyl-P-thiocytidylyl-(3′→5′)-2′-deoxy-P-thioguanylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-P-thiothymidylyl-(3′→5′)-P-thiothymidylyl-(3′→5′)-2′-deoxy-Pthiocytidylyl-(3′→5′)-P-thiothymidylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-2′-deoxy-5-methyl-P-thiocytidylyl-(3′→5′)-2′-deoxy-Pthioguanylyl-(3′→5′)-2′-deoxy-P-thiocytidylyl-(3′→5′)-2′-deoxy-Pthioadenylyl-(3′→5′)-2′-deoxy-P-thioguanylyl-(3′→5′)-2′-deoxycytidine));     -   sphingosine 1-phosphate receptor 1 (S1P1) modulators (e.g.         ozanimod         ((S)-5-(3-(1-((2-hydroxyethyl)amino)-2,3-dihydro-1H-inden-4-yl)-1,2,4-oxadiazol-5-yl)-2-isopropoxybenzonitrile),         amiselimod (MT1303;         2-amino-2-{2-[4-(heptyloxy)-3-(trifluoromethyl)phenyl]ethyl}propane-1,3-diol)         or APD334         (2-[7-[4-cyclopentyl-3-(trifluoromethyl)benzyloxy]-1,2,3,4-tetrahydrocyclopenta[b]indol-3(R)-yl]acetic         acid));     -   STAT3 inhibitors (e.g. TAK-114;         (3E)-1-methyl-3-(2-oxo-1H-indol-3-ylidene)indol-2-one);     -   Narrow spectrum kinase inhibitors (e.g., TOP1288);     -   receptor-interacting protein-1 (RIP1) kinase inhibitors (e.g.         GSK2982772);     -   Syk inhibitors and prodrugs thereof (e.g. fostamatinib and         R-406);     -   Phosphodiesterase-4 inhibitors (e.g. tetomilast);     -   HMPL-004;     -   probiotics;     -   microbiome modulators (e.g. SGM1019);     -   Dersalazine;     -   semapimod/CPSI-2364; and     -   protein kinase C inhibitors (e.g. AEB-071)

For the treatment of eye disorders (such as uveitis and keratoconjunctivitis sicca (dry eye)), the other therapeutic agent may be, for example, one or more agents selected from the list comprising:

-   -   corticosteroids (e.g. dexamethasone, prednisolone, triamcinolone         acetonide, difluprednate or fluocinolone acetonide);     -   glucocorticoid agonists (e.g. mapracorat);     -   immunosuppressants (e.g. cyclosporin, voclosporin, azathioprine,         methotrexate, mycophenolate mofetil or tacrolimus);     -   anti-TNFα antibodies (e.g. infliximab, adalimumab, certolizumab         pegol, ESBA-105 or golimumab);     -   anti-IL-17A antibodies (e.g. secukinumab);     -   mTOR inhibitors (e.g. sirolimus);     -   VGX-1027;     -   adenosine A3 receptor agonists (e.g. CF-101);     -   lifitegrast;     -   Narrow spectrum kinase inhibitors (e.g., TOP1630);     -   IL1 blockers (e.g. EBI-005; Hou et al. PNAS 2013, 110(10),         3913-3918);     -   RGN-259 (Thymosin 34);     -   SI-614;     -   OTX-101;     -   JNK inhibitors (e.g. XG-104);     -   MAP kinase signalling inhibitors (e.g. DA-6034;         {[2-(3,4-dimethoxyphenyl)-5-methoxy-4-oxochromen-7-yl]oxy}acetic         acid);     -   mucin stimulators (e.g. rebamipide;         2-[(4-chlorobenzoyl)amino]-3-(2-oxo-1H-quinolin-4-yl)propanoic         acid);     -   MIM-D3 (Tavilermide; see, for example, US 2013/0345395); and     -   protein kinase C inhibitors (e.g. AEB-071).

Medical Uses

The compounds of the invention may be used as monotherapies for inflammatory diseases, or in combination therapies for such diseases.

Thus, embodiments of aspects (e) to (g) above that may be mentioned include those in which the compound of formula I, Ia, Ib or Ic (or pharmaceutically acceptable salt, solvate or isotopic derivative thereof) is the sole pharmacologically active ingredient utilised in the treatment.

However, in other embodiments of aspects (e) to (g) above, the compound of formula I, Ia, Ib or Ic (or pharmaceutically acceptable salt, solvate or isotopic derivative thereof) is administered to a subject who is also administered one or more other therapeutic agents (e.g. wherein the one or more other therapeutic agents are as defined above in connection with combination products).

When used herein, the term “inflammatory disease” specifically includes references to any one or more of the following:

-   (i) lung diseases or disorders having an inflammatory component,     such as cystic fibrosis, pulmonary hypertension, lung sarcoidosis,     idiopathic pulmonary fibrosis or, particularly, COPD (including     chronic bronchitis and emphysema), asthma or paediatric asthma; -   (ii) skin diseases or disorders having an inflammatory component,     such as atopic dermatitis, allergic dermatitis, contact dermatitis     or psoriasis; -   (iii) nasal diseases or disorders having an inflammatory component,     such as allergic rhinitis, rhinitis or sinusitis; -   (iv) eye diseases or disorders having an inflammatory component,     such as conjunctivitis, allergic conjunctivitis, glaucoma, diabetic     retinopathy, macular oedema (including diabetic macular oedema),     central retinal vein occlusion (CRVO), dry and/or wet age related     macular degeneration (AMD), post-operative cataract inflammation,     or, particularly, keratoconjunctivitis sicca (dry eye, also known as     xerophthalmia), uveitis (including posterior, anterior and pan     uveitis), corneal graft and limbal cell transplant rejection; and -   (v) gastrointestinal diseases or disorders having an inflammatory     component, such as gluten sensitive enteropathy (coeliac disease),     eosinophilic esophagitis, intestinal graft versus host disease or,     particularly, Crohn's disease or ulcerative colitis.

References herein to diseases having an inflammatory component include references to diseases that involve inflammation, whether or not there are other (non-inflammatory) symptoms or consequences of the disease.

According to a further aspect of the invention there is provided a process for the preparation of a compound of formula I, which process comprises:

(a) reaction of a compound of formula II,

wherein X, Ak, s, L and Q are as hereinbefore defined and LG represents a leaving group (e.g., halogen, methanesulfonyl or triflate), with a compound of formula III,

wherein R¹ and R² are as hereinbefore defined, for example under conditions known to those skilled in the art, such as through (i) a typical S_(N)Ar reaction in the presence of a suitable organic solvent (e.g., a polar aprotic solvent such as DMSO, DMF, THF, or mixtures thereof) and a base (e.g., DIPEA or TEA) or (ii) via a Buchwald-Hartwig coupling (Lundgren, R. J.; Stradiotto, M. Aldrichim. Acta 2012, 45(3), 59-65) employing a transition metal catalyst, e.g., palladium, and a suitable ligand, e.g., BrettPhos (J. Am. Chem. Soc. 2008, 130, 13552-13554); (b) for compounds of formula I in which L represents C(O), reaction of a compound of formula IV,

wherein X, Ak and s are as hereinbefore defined, with a compound of formula VIII,

wherein Q, R¹ and R² are as hereinbefore defined, for example under conditions known to those skilled in the art, such as reaction with the compound of formula VIII under standard amide bond forming conditions (e.g., involving a coupling agent, such as T3P or HATU (WO2013/014567), optionally in the presence of a base, such as DIPEA, in a polar aprotic solvent, such as DMF); (c) reaction of a compound of formula IV, as hereinbefore defined, with a compound of formula IX,

wherein LG¹ represents a leaving group (e.g., halogen or sulfonate) and L, Q, R¹ and R² are as hereinbefore defined, for example under conditions known to those skilled in the art, such as in the presence of a base, such as DIPEA, and a suitable inert solvent (e.g., DMF), optionally at elevated temperatures of up to 50° C.; (d) for compounds of formula I in which L represents CH₂, reaction of a compound of formula IV, as hereinbefore defined, with a compound of formula X,

wherein Q, R¹ and R² are as hereinbefore defined, for example under conditions known to those skilled in the art, such as standard reductive alkylation conditions in the presence of a reducing agent (e.g., sodium triacetoxyborohydride: J. Am. Chem. Soc. 1996, 61, 3849-3862) and a suitable inert solvent (e.g., 1,2-dichloroethane, THF or DMF), optionally in the presence of acetic acid; or (e) deprotection of an protected derivative of a compound of formula I, under conditions known to those skilled in the art, wherein the protected derivative bears a protecting group on an O- or N-atom of the compound of formula I (and, for the avoidance of doubt, a protected derivative of one compound of formula I may or may not represent another compound of formula I).

Examples of protected derivatives of compounds of formula I include those where:

-   -   an O-atom is protected with a benzyl group, which benzyl group         may be removed by hydrogenation, for example in the presence of         a palladium catalyst (such as Pd/C);     -   an O-atom of an acid (e.g. a carboxylic acid) is protected with         an alkyl group (such as methyl, ethyl or tert-butyl), which         alkyl group may be removed by either basic hydrolysis (e.g. for         methyl or ethyl groups, by a hydrolysis reaction using an alkali         metal hydroxide such as sodium hydroxide) or acid hydrolysis         (e.g. for a tert-butyl group, by a hydrolysis reaction using an         acid such as trifluoroacetic acid);     -   an N-atom of an amine is protected with a carbamate group, such         as a benzyl or tert-butyl carbamate, which groups may be removed         under similar conditions to those used to remove benzyl or         tert-butyl groups from O-atoms.

The compound of formula II can be prepared according to procedures similar to those outlined in WO2013/014567. Thus, the compound of formula II can be produced (Scheme 1) by condensing an amine of formula IV with (i) an acid of formula V under standard amide bond forming conditions (e.g., involving a coupling agent, such as T3P or HATU, optionally in the presence of a base, such as DIPEA, in a polar aprotic solvent, such as DMF), (ii) an electrophile of formula VI, i.e., an acid chloride, sulfonyl chloride, alkyl halide or activated aryl halide, where LG¹ represents a leaving group (e.g., halogen or sulfonate) displaced preferentially before LG, in the presence of a base, such as DIPEA, and a suitable inert solvent (e.g., DMF), optionally at elevated temperatures of up to 50° C., or (iii) an aldehyde of formula VII in the presence of a reducing agent (e.g., sodium triacetoxyborohydride: J. Am. Chem. Soc. 1996, 61, 3849-3862) and a suitable inert solvent (e.g., 1,2-dichloroethane, THF or DMF), optionally in the presence of acetic acid.

Compounds of formula VIII, IX and X may be prepared by reacting a compound of formula III, as hereinbefore defined, with a compound of formula V, VI or VII, respectively, through either S_(N)Ar or Buchwald-Hartwig couplings, as outlined above.

Compounds of formula III, V, VI and VII are either commercially-available, or can be obtained using the procedures cited, or can be readily prepared by conventional methods by those skilled in the art.

The aspects of the invention described herein (e.g. the above-mentioned compounds, combinations, methods and uses) may have the advantage that, in the treatment of the conditions described herein, they may be more convenient for the physician and/or patient than, be more efficacious than, be less toxic than, have better selectivity over, have a broader range of activity than, be more potent than, produce fewer side effects than, have a better pharmacokinetic and/or pharmacodynamic profile than, have more suitable solid state morphology than, have better long term stability than, or may have other useful pharmacological properties over similar compounds, combinations, methods (treatments) or uses known in the prior art for use in the treatment of those conditions or otherwise.

The compounds of the invention may additionally (or alternatively):

-   -   exhibit a long duration of action and/or persistence of action         (e.g. in comparison to the Reference Compound);     -   be more potent in biochemical JAK enzyme assays (e.g. in         comparison to the Reference Compound);     -   exhibit lower IC₅₀ values in cellular assays evaluating cytokine         release (e.g. in comparison to the Reference Compound);     -   produce lower systemic concentrations following oral dosing         (e.g. in comparison to the Reference Compound);     -   maintain a relatively high local drug concentration between         doses (e.g. a high local concentration relative to other         previously disclosed JAK inhibitors);     -   exhibit properties that are particularly suited to topical/local         administration (e.g. following topical/local administration, the         generation of high target tissue concentrations but low plasma         concentrations of the compounds of formula (I) and/or rapid         clearance of the compounds of formula (I) from plasma, for         example as a result of high renal or hepatic extraction);     -   display reduced cytotoxicities (e.g. in comparison to the         Reference Compound); and/or     -   exhibit enhanced solubilities (e.g. in comparison to the         Reference Compound) in biologically-relevant media, for example,         in fasted-state simulated colonic fluid (FaSSCoF; Vertzoni, M.,         et al. Pharm. Res. 2010, 27, 2187-2196).

EXPERIMENTAL SECTION

Abbreviations used herein are defined below. Any abbreviations not defined are intended to convey their generally accepted meaning.

CHEMISTRY EXAMPLES

General Procedures

All starting materials and solvents were obtained either from commercial sources or prepared according to the literature citation. Unless otherwise stated, all reactions were stirred. Organic solutions were routinely dried over anhydrous magnesium sulfate.

Preparative Reverse Phase High Performance Liquid Chromatography

Performed using UV detection at 210-400 nm with a Waters X-Bridge Prep-C18, 5 μm, 19×50 mm column eluting with a H2O-MeCN gradient containing 0.1% ammonium bicarbonate over 10 min.

Analytical Methods

Reverse Phase High Performance Liquid Chromatography

Method 1:

Agilent Infinity, X-Select, Waters X-Select C18, 2.5 μm (4.6×30 mm) at 40° C.; flow rate 2.5-4.5 mL min⁻¹ eluted with a H₂O-MeCN gradient containing 0.1% v/v formic acid over 4 min employing UV detection at 254 nm. Gradient information: 0-3.00 min, ramped from 95% H₂O-5% MeCN to 5% H₂O-95% MeCN; 3.00-3.01 min, held at 5% H₂O-95% MeCN, flow rate increased to 4.5 mL min⁻¹; 3.01-3.50 min, held at 5% H₂O-95% MeCN; 3.50-3.60 min, returned to 95% H₂O-5% MeCN, flow rate reduced to 3.50 mL min⁻¹; 3.60-3.90 min, held at 95% H₂O-5% MeCN; 3.90-4.00 min, held at 95% H₂O-5% MeCN, flow rate reduced to 2.5 mL min⁻¹.

Method 2:

Waters Acquity UPLC C18, 1.7 μm (2.1×50 mm) at 40° C.; inject volume 2 μL; flow rate 0.77 mL min⁻¹ eluted with a H₂O-MeCN gradient containing 10 mM ammonium bicarbonate over 3 min employing UV detection at 210-400 nm. Gradient information: 0-0.11 min, held at 95% H₂O-5% MeCN; 0.11-2.15 min, ramped from 95% H₂O-5% MeCN to 5% H₂O-95% MeCN; 2.15-2.49 min, held at 5% H₂O-95% MeCN; 2.49-2.56 min, returned to 95% H₂O-5% MeCN; 2.56-3.00 min, held at 95% H₂O-5% MeCN.

¹H NMR Spectroscopy

¹H NMR spectra were acquired on a Bruker Avance III spectrometer at 400 MHz using residual undeuterated solvent as reference and unless specified otherwise were run in DMSO-d₆.

Preparation of Compounds of the Invention Example 1 (2-(6-(2-Ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(methyl(1-methylpiperidin-4-yl)amino)pyrazin-2-yl)methanone

(i) Intermediate A: (5-Chloropyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone

To a stirred solution of 5-ethyl-2-fluoro-4-(3-(4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridin-2-yl)-1H-indazol-6-yl)phenol.2HCl (Coe, J. W., et al. WO2013/014567, 31 Jan. 2013; 500 mg, 1.11 mmol), 5-chloropyrazine-2-carboxylic acid (176 mg, 1.11 mmol) and DIPEA (970 μL, 5.55 mmol) in DMF (10 mL) was added T₃P (50% in EtOAc) (1.90 mL, 3.33 mmol) and the resulting yellow solution left to stir at rt for 2 h. The reaction mixture was diluted with water (10 mL) to form a precipitate. The precipitate was collected by filtration and then washed with water (10 mL) and diethyl ether (10 mL) to give a pale yellow solid. The solid was dissolved in MeOH (15 mL) and concentrated in vacuo to afford the sub-title compound as a yellow solid (521 mg).

¹H NMR (400 MHz, DMSO-d₆) δ: 13.30-13.19 (m, 1H), 12.58 (s, 1H), 8.88 (m, 1H), 8.78 (m, 1H), 8.34 and 8.25 (2×d, 1H), 7.39 and 7.37 (2×s, 1H), 7.15-6.98 (m, 2H), 6.96-6.87 (m, 1H), 4.76 and 4.59 (2×s, 2H), 4.04 (t, 1H), 3.73 (t, 1H), 2.89 (s, 1H), 2.84-2.71 (m, 2H), 2.49-2.43 (m, 1H), 1.30-1.20 (m, 1H), 1.01 (m, 3H).

LCMS m/z 518 (M+H)⁺ (ES⁺)

(ii) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(methyl(1-methylpiperidin-4-yl)amino)pyrazin-2-yl)methanone

A solution of the product from step (i) (Intermediate A; 75.0 mg, 0.145 mmol) in DMSO (2 mL) was added to N,1-dimethylpiperidin-4-amine (4.35 mg, 0.290 mmol), followed by DIPEA (101 μL, 0.579 mmol). The solution was stirred at rt for 18 h. The reaction mixture was filtered and then purified by preparative HPLC (Waters, Basic (0.1% Ammonium Bicarbonate), Basic, Waters X-Bridge Prep-C18, 5 μm, 19×50 mm column, 10-40% MeCN in Water) to afford the title compound as a white solid (16.3 mg).

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), 12.52 and 12.27 (2×s, 1H), 9.86 (s, 1H), 8.43-8.38 (m, 1H), 8.37-8.21 (m, 1H), 8.16 (s, 1H), 7.37 (s, 1H), 7.16-6.97 (m, 2H), 6.91 (d, 1H), 4.83-4.62 (m, 2H), 4.48-4.36 (m, 1H), 4.01-3.86 (m, 2H), 3.30 (s, 2H), 2.99 (s, 3H), 2.90-2.73 (m, 4H), 2.18 (s, 3H), 2.02 (t, 2H), 1.90-1.75 (m, 2H), 1.57 (d, 2H), 1.02 (t, 3H).

LCMS m/z 610.5 (M+H)⁺ (ES⁺)

Example 2

The following compounds were prepared by reacting the appropriate amine with Intermediate A employing methods analogous to those described above for Example 1 (e.g. at temperatures of 20-90° C.). The only exception was Example 2(a), which was made by reacting Intermediate A with methyl 4-(methylamino)butanoate, then the resulting methyl ester was isolated and saponified with LiOH, before being purified by RP-HPLC, to afford the corresponding acid.

(a) 4-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)butanoic acid

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), 12.51 (s, 1H), 9.95 (s, 1H), 8.47-8.23 (m, 2H), 8.17 (s, 1H), 7.37 (s, 1H), 7.16-6.97 (m, 2H), 6.92 (d, 1H), 4.91-4.60 (m, 2H), 3.94 (s, 3H), 3.60 (t, 2H), 3.12 (d, 3H), 2.79 (s, 4H), 2.25-2.17 (m, 2H), 1.84-1.72 (m, 2H), 1.02 (t, 3H).

LCMS m/z 599.5 (M+H)⁺ (ES⁺)

(b) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(4-hydroxypiperidin-1-yl)ethyl)amino)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), 12.52 and 12.26 (2×s, 1H), 9.81 (s, 1H), 8.43-8.20 (m, 2H), 7.95 (d, 1H), 7.51 (s, 1H), 7.37 (s, 1H), 7.17-6.97 (m, 2H), 6.91 (d, 1H), 4.86-4.59 (m, 2H), 4.54 (t, 1H), 3.91 (s, 2H), 3.49-3.37 (m, 3H), 2.85-2.69 (m, 4H), 2.49-2.41 (m, 4H), 2.06 (t, 2H), 1.76-1.63 (m, 2H), 1.45-1.31 (m, 2H), 1.02 (t, 3H).

LCMS m/z 626.5 (M+H)⁺ (ES⁺)

(c) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(methyl(pyridin-2-ylmethyl)amino)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), 12.53 and 12.28 (2×s, 1H), 9.79 (s, 1H), 8.57-8.49 (m, 1H), 8.45-8.17 (m, 3H), 7.81-7.71 (m, 1H), 7.38 (s, 1H), 7.33-7.23 (m, 2H), 7.16-6.99 (m, 2H), 6.92 (d, 1H), 4.96 (s, 2H), 4.84-4.62 (m, 2H), 3.93 (s, 2H), 3.26 (s, 3H), 2.79 (s, 2H), 2.50-2.44 (m, 2H), 1.02 (t, 3H).

LCMS m/z 604.5 (M+H)⁺ (ES⁺)

(d) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(1-oxidothiomorpholino)ethyl)amino)pyrazin-2-yl)methanone

¹H NMR (400 MHz, Methanol-d₄) δ: 8.39 (d, 1H), 8.28 (s, 1H), 7.96 (d, 1H), 7.41 (s, 1H), 7.16 (s, 1H), 6.92 (dd, 2H), 4.95-4.89 (assume 2H, obscured by solvent), 4.06 (s, 2H), 3.59 (t, 2H), 3.38-3.34 (assume 4H, obscured by solvent), 3.18-2.81 (m, 10H), 2.73 (t, 2H), 2.55 (q, 2H), 1.07 (t, 3H).

LCMS m/z 644.5 (M+H)⁺ (ES⁺)

(e) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(1-oxildothiomorpholino)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.21 (s, 1H), 12.52 and 12.29 (2×s, 1H), 9.80 (s, 1H), 8.52-8.41 (m, 2H), 8.41-8.19 (m, 1H), 7.38 (s, 1H), 7.18-6.98 (m, 2H), 6.91 (d, 1H), 4.91-4.59 (m, 2H), 4.44-4.25 (m, 2H), 4.08-3.82 (m, 4H), 2.97 (ddd, 2H), 2.88-2.70 (m, 4H), 2.49-2.43 (assume 2H, obscured by solvent), 1.02 (t, 3H).

LCMS m/z 601.4 (M+H)⁺ (ES⁺)

(f) (5-(4-(2-(dimethylamino)ethyl) piperazin-1-yl)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.21 (s, 1H), 12.51 and 12.26 (2×s, 1H), 9.85 (s, 1H), 8.45-8.20 (m, 3H), 7.37 (s, 1H), 7.18-6.96 (m, 2H), 6.91 (d, 1H), 4.83-4.60 (m, 2H), 4.03-3.82 (m, 2H), 3.72-3.59 (m, 4H), 2.78 (s, 2H), 2.54-2.51 (assume 4H, obscured by solvent), 2.48-2.34 (m, 6H), 2.15 (s, 6H), 1.01 (t, 3H).

LCMS m/z 639.6 (M+H)⁺ (ES⁺)

(g) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(4-hydroxy-1-methylpiperidin-4-yl)ethyl)amino)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), 12.52 and 12.27 (2×s, 1H), 9.68 (s, 1H), 8.43-8.21 (m, 2H), 7.89 (d, 1H), 7.53 (s, 1H), 7.38 (s, 1H), 7.17-6.97 (m, 2H), 6.92 (d, 1H), 4.85-4.59 (m, 2H), 4.18 (d, 1H), 3.92 (s, 2H), 3.46-3.36 (m, 2H), 2.86-2.71 (m, 2H), 2.49-2.43 (assume 2H, obscured by solvent), 2.42-2.20 (m, 4H), 2.14 (s, 3H), 1.74-1.62 (m, 2H), 1.58-1.44 (m, 4H), 1.02 (t, 3H).

LCMS m/z 640.5 (M+H)⁺ (ES⁺)

(h) (S)-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(2-(hydroxymethyl)pyrrolidin-1-yl)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-de) δ: 13.22 (s, 1H), 12.52 and 12.29 (2×s, 1H), 9.83 (s, 1H), 8.49-8.20 (m, 2H), 8.04 (s, 1H), 7.38 (s, 1H), 7.20-6.96 (m, 2H), 6.92 (d, 1H), 4.95-4.58 (m, 3H), 4.24-4.11 (m, 1H), 4.08-3.86 (m, 2H), 3.67-3.51 (m, 2H), 3.46-3.36 (assume 2H, obscured by solvent), 2.80 (s, 2H), 2.49-2.41 (assume 2H, obscured by solvent), 2.13-1.86 (m, 4H), 1.02 (t, 3H).

LCMS m/z 583.5 (M+H)⁺ (ES⁺)

(i) (5-(bis(2-methoxyethyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), 12.52 and 12.28 (2×s, 1H), 9.83 (s, 1H), 8.44-8.12 (m, 3H), 7.37 (s, 1H), 7.19-6.96 (m, 2H), 6.91 (d, 1H), 4.88-4.55 (m, 2H), 4.05-3.84 (m, 2H), 3.78 (t, 4H), 3.55 (t, 4H), 3.26 (s, 6H), 2.79 (s, 2H), 2.49-2.43 (assume 2H, obscured by solvent), 1.01 (t, 3H).

LCMS m/z 615.5 (M+H)⁺ (ES⁺)

(j) (5-(4-(2-(1H-imidazol-1-yl)ethyl)piperazin-1-yl)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), 12.53 and 12.28 (2×s, 1H), 9.87 (s, 1H), 8.49-8.19 (m, 3H), 7.66 (s, 1H), 7.38 (s, 1H), 7.21 (s, 1H), 7.18-6.97 (m, 2H), 6.97-6.83 (m, 2H), 4.87-4.57 (m, 2H), 4.13 (t, 2H), 4.05-3.82 (m, 2H), 3.73-3.61 (m, 4H), 2.79 (s, 2H), 2.69 (t, 2H), 2.60-2.53 (m, 4H), 2.50-2.42 (assume 2H, obscured by solvent), 1.02 (t, 3H).

LCMS m/z 662.5 (M+H)⁺ (ES⁺)

(k) 1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl) pyrazin-2-yl)(methyl)amino)ethyl)pyrrolidin-2-one

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), 12.52 and 12.31 (2×s, 1H), 9.83 (s, 1H), 8.46-8.21 (m, 2H), 8.12 (s, 1H), 7.38 (s, 1H), 7.23-6.97 (m, 2H), 6.92 (d, 1H), 4.93-4.60 (m, 2H), 4.07-3.86 (m, 2H), 3.79 (t, 2H), 3.46-3.37 (m, 4H), 3.12 (s, 3H), 2.80 (s, 2H), 2.48-2.44 (assume 2H, obscured by solvent), 2.05 (t, 2H), 1.88-1.73 (m, 2H), 1.02 (t, 3H).

LCMS m/z 624.6 (M+H)⁺ (ES⁺)

(l) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-(4-(2-hydroxyethyl)piperazin-1-yl)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.28, 1H), 9.87 (s, 1H), 8.45-8.38 (m, 1H), 8.37-8.22 (m, 2H), 7.41-7.34 (m, 1H), 7.15-6.98 (m, 2H), 6.91 (d, 1H), 4.81-4.61 (m, 2H), 4.47 (t, 2H), 4.02-3.80 (m, 2H), 3.70-3.61 (m, 4H), 3.55 (q, 2H), 2.84-2.72 (m, 2H), 1.01 (t, 3H). 7H under DMSO peak.

LCMS m/z 612 (M+H)⁺ (ES⁺)

(m) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino) pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.28, 1H), 9.87 (s, 1H), 8.45-8.38 (m, 1H), 8.37-8.22 (m, 2H), 7.41-7.34 (m, 1H), 7.15-6.98 (m, 2H), 6.91 (d, 1H), 4.81-4.61 (m, 2H), 4.47 (t, 2H), 4.02-3.80 (m, 2H), 3.70-3.61 (m, 4H), 3.55 (q, 2H), 2.84-2.72 (m, 2H), 1.01 (t, 3H). 7H under DMSO peak.

LCMS m/z 645 (M+H)⁺ (ES⁺)

(n) N-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methy)amino)ethyl)-N-methylacetamide

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.31, 1H), 9.88 (s, 1H), 8.40 (dd, 1H), 8.38-8.17 (m, 1H), 8.15 (d, 1H), 7.37 (s, 1H), 7.10 (d, 1H), 7.03 (d, 1H), 6.91 (d, 1H), 4.82-4.66 (m, 2H), 3.99-3.86 (m, 2H), 3.76 (dt, 2H), 3.50 (q, 2H), (2×s, 3.17 and 3.11, 3H), 2.97 (s, 2H), 2.84 (s, 1H), 2.84-2.70 (m, 2H), (2×s, 1.92 and 1.96, 3H), 1.01 (t, 3H). 2H under DMSO peak.

LCMS m/z 612 (M+H)⁺ (ES⁺)

(o) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-(3-(hydroxymethyl)azetidin-1-yl)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.54 and 12.29, 1H), 9.87 (s, 1H), 8.40-8.22 (m, 2H), 7.86 (s, 1H), 7.38 (s, 1H), 7.11 (d, 1H), 7.03 (d, 1H), 6.92 (d, 1H), 4.87-4.79 (m, 1H), 4.74-4.66 (m, 2H), 4.14 (t, 2H), 3.96 (s, 1H), 3.88 (t, 3H), 3.61 (t, 2H), 2.96-2.73 (m, 3H), 1.02 (t, 3H). 2H under DMSO peak.

LCMS m/z 569 (M+H)⁺ (ES⁺)

(p) (5-(bis(2-hydroxyethyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.28, 1H), 9.89 (s, 1H), 8.42-8.27 (m, 2H), 8.20 (s, 1H), 7.37 (s, 1H), 7.10 (d, 1H), 7.02 (d, 1H), 6.92 (d, 1H), 4.87-4.68 (m, 2H), 4.77-4.58 (m, 2H), 4.00-3.86 (m, 2H), 3.73-3.57 (m, 7H), 2.86-2.70 (m, 2H), 1.01 (t, 3H). 3H under DMSO peak.

LCMS m/z 587 (M+H)⁺ (ES⁺)

(q) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2-(methylsulfonyl)ethyl)amino)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.27, 1H), 9.88 (s, 1H), 8.81-8.19 (m, 2H), 7.98 (s, 1H), 7.94-7.85 (m, 1H), 7.38 (s, 1H), 7.11 (d, 1H), 7.02 (d, 1H), 6.92 (d, 1H), 4.84-4.60 (m, 2H), 4.00-3.83 (m, 2H), 3.76 (q, 2H), 3.41 (t, 2H), 3.05 (s, 3H), 2.85-2.61 (m, 2H), 1.02 (t, 3H). 2H under DMSO peak.

LCMS m/z 605 (M+H)⁺ (ES⁺)

(r) 1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)amino)ethyl)pyrrolidin-2-one

¹H NMR (400 MHz, DMSO-d₆) δ: 13.22 (s, 1H), (2×s, 12.53 and 12.27, 1H), 9.87 (s, 1H), 8.38-8.25 (m, 2H), 7.89 (s, 1H), 7.75-7.52 (m, 1H), 7.37 (s, 1H), 7.10 (d, 1H), 7.03 (d, 1H), 6.92 (d, 1H), 4.81-4.57 (m, 2H), 3.99-3.80 (m, 2H), 3.52-3.43 (m, 2H), 3.42-3.33 (m, 4H), 23.75-2.71 (m, 2H), 2.17 (t, 2H), 1.89 (p, 2H), 1.01 (t, 3H). 2H under DMSO peak.

LCMS m/z 610 (M+H)⁺ (ES⁺)

(s) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidin-1-yl)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.27, 1H), 9.88 (s, 1H), 8.41-8.22 (m, 3H), 7.38 (s, 1H), 7.18-7.07 (m, 1H), 7.01 (d, 1H), 6.91 (d, 1H), 5.42-4.93 (m, 3H), 4.91-4.58 (m, 3H), 4.39-4.32 (m, 1H), 4.27 (d, 1H), 4.03-3.87 (m, 2H), 3.83-3.64 (m, 4H), 3.56-3.51 (m, 1H), 3.43 (dd, 1H), 2.87-2.72 (m, 2H), 1.02 (t, 3H). 2H under DMSO peak.

LCMS m/z 645 (M+H)⁺ (ES⁺)

(t) (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)pyrazin-2-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.24 (s, 1H), (2×s, 12.53 and 12.29, 1H), 9.91 (s, 1H), 8.49-8.27 (m, 2H), 8.00 (s, 1H), 7.38 (s, 1H), 7.11 (d, 1H), 7.03 (d, 1H), 6.92 (d, 1H), 4.81-4.67 (m, 2H), 4.00-3.84 (m, 2H), 3.76-3.71 (m, 2H), 3.44 (dd, 2H), 3.02-2.89 (m, 2H), 2.85-2.72 (m, 2H), 2.22 (s, 3H), 1.02 (t, 3H). 2H under water peak. 2H under DMSO peak.

LCMS m/z 608 (M+H)⁺ (ES⁺)

(u) 3-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)ethyl)oxazolidin-2-one

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.54 and 12.30, 1H), 9.87 (s, 1H), 8.45-8.40 (m, 1H), 8.37-8.24 (m, 1H), 8.15 (s, 1H), 7.38 (s, 1H), 7.07 (dd, 2H), 6.92 (d, 1H), 4.85-4.64 (m, 2H), 4.15 (t, 2H), 4.02-3.89 (m, 2H), 3.82 (t, 2H), 3.63 (t, 2H), 3.42 (t, 2H), 3.14 (s, 3H), 2.88-2.74 (m, 2H), 1.02 (t, 3H). 2H under DMSO peak.

LCMS m/z 626 (M+H)⁺ (ES⁺)

(v) (5-((2-(1,1-dioxidoisothiazolidin-2-yl)ethyl)(methyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)methanone

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.30, 1H), 9.88 (s, 1H), 8.45-8.39 (m, 1H), 8.36-8.22 (m, 1H), 8.16 (s, 1H), 7.38 (s, 1H), 7.11 (d, 1H), 7.02 (d, 1H), 6.92 (d, 1H), 4.83-4.67 (m, 2H), 4.01-3.87 (m, 2H), 3.81 (t, 2H), 3.30 (t, 2H), 3.20-3.11 (m, 7H), 2.85-2.73 (m, 2H), 2.18 (p, 2H), 1.02 (t, 3H). 2H under DMSO peak.

LCMS m/z 660 (M+H)⁺ (ES⁺)

(w) (3aR,6aS)-5-(5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)tetrahydropyrrolo[3,4-c]pyrrole-1,3(2H,3aH)-dione

¹H NMR (400 MHz, DMSO-d₆) δ: 13.23 (s, 1H), (2×s, 12.53 and 12.26, 1H), 11.41 (s, 1H), 9.87 (s, 1H), 8.44 (s, 1H), 8.38-8.23 (m, 1H), 8.18 (s, 1H), 7.43-7.35 (m, 1H), 7.15-6.99 (m, 2H), 6.92 (d, 1H), 4.83-4.64 (m, 2H), 4.07 (d, 2H), 4.00-3.83 (m, 2H), 3.69-3.55 (m, 4H), 2.83-2.61 (m, 2H), 1.02 (t, 3H). 2H under DMSO peak.

LCMS m/z 622 (M+H)⁺ (ES⁺)

Biological Testing: Experimental Methods

Enzyme Binding Assays (Kinomescan)

Kinase enzyme binding activities of compounds disclosed herein may be determined using a proprietary assay which measures active site-directed competition binding to an immobilized ligand (Fabian, M. A., et al., Nature Biotechnol. 2005, 23, 329-336). These assays may be conducted by DiscoveRx (formerly Ambit; San Diego, Calif.). The percentage inhibition produced by incubation with a test compound may be calculated relative to the non-inhibited control.

Cellular Assays

The compounds of the invention were studied using one or more of the following assays.

(a) IFNγ Release from CD3/IL2 Stimulated PBMC Cells PBMCs from healthy subjects are separated from whole blood using a density gradient (Lymphoprep, Axis-Shield Healthcare). Cells are added to a 96 well plate, and compound at the desired concentration added 2 hours prior to stimulation with a mixture of monoclonal antibody to CD3 (1 μg/ml, eBioscience) and human recombinant IL2 (10 ng/ml, Peprotech). After 48 hours incubation under normal tissue culture conditions, supernatants are collected and IFNγ release determined by Sandwich ELISA (Duo-set, R&D System). The IC₅₀ is determined from the dose response curve.

(b) IL-2 and IFNγ Release in CD3/CD28 Stimulated LPMC Cells from IBD Patients

Lamina propria mononuclear cells (LPMCs) are isolated and purified from inflamed IBD mucosa of surgical specimens or from normal mucosa of surgical specimens as follows: The mucosa is removed from the deeper layers of the surgical specimens with a scalpel and cut in fragments of size 3-4 mm. The epithelium is removed by washing the tissue fragments three times with 1 mM EDTA (Sigma-Aldrich, Poole, UK) in HBSS (Sigma-Aldrich) with agitation using a magnetic stirrer, discarding the supernatant after each wash. The sample is subsequently treated with type 1A collagenase (1 mg/mL; Sigma-Aldrich) for 1 h with stirring at 37° C. The resulting cell suspension is then filtered using a 100 μm cell strainer, washed twice, resuspended in RPMI-1640 medium (Sigma-Aldrich) containing 10% fetal calf serum, 100 U/mL penicillin and 100 μg/mL streptomycin, and used for cell culture.

Freshly isolated LPMCs (2×10⁵ cells/well) are stimulated with 1 μg/mL α-CD3/α-CD28 for 48 h in the presence of either DMSO control or appropriate concentrations of compound. After 48 h, the supernatant is removed and assayed for the presence of TNFα and IFNγ by R&D ELISA. Percentage inhibition of cytokine release by the test compounds is calculated relative to the cytokine release determined for the DMSO control (100%).

(c) Cell Cytotoxicity Assay

1×10⁵ Jurkat cells (immortalised human T lymphocytes) are added to the appropriate number of wells of a 96 well plate in 100 μL of media (RPMI supplemented with 10% foetal bovine serum). 1 μL of DMSO control (final concentration 1.0% v/v) or test compound (final concentration 20, 5 or 1 μg/mL) is added to the wells and incubated at 37° C., 5% CO₂. After 24 hours, the plate is centrifuged at 1200 rpm for 3 minutes and the supernatant discarded. Cells are then resuspended in 150 μL (final concentration 7.5 μg/mL) of propidium iodide (PI) in PBS and incubated at 37° C., 5% CO₂ for 15 minutes. After 15 minutes, cells are analysed by flow cytometry (BD accuri) using the FL3 window. The % viability is calculated as the % of cells that are PI negative in the test wells normalised to the DMSO control.

In Vivo Screening: Pharmacodynamics and Anti-Inflammatory Activity

(i) DSS-Induced Colitis in Mice

Non-fasted, 10-12 week old, male BDF1 mice are dosed by oral gavage twice daily with either vehicle, reference item (5-ASA) or test compound one day before (Day −1) stimulation of the inflammatory response by treatment with dextran sodium sulphate (DSS). On Day 0 of the study, DSS (5% w/v) is administered in the drinking water followed by BID dosing of the vehicle (5 mL/kg), reference (100 mg/kg) or test compound (5 mg/kg) for 7 days. The drinking water with DSS is replenished every 3 days. During the study, animals are weighed every day and stool observations are made and recorded as a score, based on stool consistency. At the time of sacrifice on Day +6, the large intestine is removed and the length and weight are recorded. Sections of the colon are taken for either MPO analysis, to determine neutrophil infiltration, or for histopathology scoring to determine disease severity.

(ii) TNBS-Induced Colitis in Mice

Non-fasted, 10-12 week old, male BDF1 mice are dosed by oral gavage twice daily with either vehicle (5 mL/kg), reference item (Budesonide 2.5 mg/kg) or test compound (1 or 5 mg/kg) one day before (Day −1) stimulation of the inflammatory response by treatment with 2,4,6-trinitrobenzenesulphonic acid (TNBS) (15 mg/mL in 50% ethanol/50% saline). On Day 0 of the study TNBS (200 μL) is administered intra-colonically via a plastic catheter with BID dosing of the vehicle, reference or test compound continuing for 2 or 4 days. During the study, animals are weighed every day and stool observations are made and recorded as a score, based on stool consistency. At the time of sacrifice on Day 2 (or Day 4), the large intestine is removed and the length and weight recorded. Sections of the colon are taken for histopathology scoring to determine disease severity.

(iii) Adoptive Transfer in Mice

On Study day 0, female Balb/C mice are terminated and spleens obtained for CD45RB^(high) cell isolation (Using SCID IBD cell Separation protocol). Approximately 4×10⁵ cells/mL CD45RB^(high) cells are then injected intraperitoneally (100 μL/mouse) into female SCID animals. On study day 14, mice are weighed and randomized into treatment groups based on body weight. On Day 14, compounds are administered BID, via oral gavage, in a dose volume of 5 mL/kg. Treatment continues until study day 49, at which point the animals are necropsied 4 hours after the morning administration. The colon length and weight are recorded and used as a secondary endpoint in the study as a measurement of colon oedema. The colon is then divided into six cross-sections, four of which are used for histopathology scoring (primary endpoint) and two are homogenised for cytokine analysis. Data is given as the % inhibition of the induction window between naïve animals and vehicle animals, where higher inhibition implies closer to the non-diseased, naïve, phenotype.

(iv) Endotoxin-Induced Uveitis in Rats

Male, Lewis rats (6-8 weeks old, Charles River UK Limited) are housed in cages of 3 at 19-21° C. with a 12 h light/dark cycle (07:00/19:00) and fed a standard diet of rodent chow and water ad libitum. Non-fasted rats are weighed, individually identified on the tail with a permanent marker, and receive a single intravitreal administration into the right vitreous humor (5 μL dose volume) of 100 ng/animal of LPS (Escherichia coli 0111:B4 prepared in PBS, Sigma Aldrich, UK) using a 32-gauge needle. Untreated rats are injected with PBS. Test compound or vehicle (4% polyoxyl 40 stearate, 4% mannitol in PBS (pH 7.4)) are administered by the topical route onto the right eye (10 μL) of animals 1 hour prior to LPS, at the time of LPS administration, and 1, 2 and 4 hours post LPS administration. Before administration, the solution to be administered is sonicated to ensure a clear solution. 6 hours after LPS dosing, animals are euthanized by overdose with pentobarbitone (via cardiac puncture). Immediately after euthanasia, 10 μL of aqueous humor is collected from the right eye of the rats by puncture of the anterior chamber using a 32 gauge needle under a surgical microscope. The aqueous humor is diluted in 20 μL of PBS and total cell counts are measured immediately using a Countess automated cell counter (Invitrogen). Following collection of the aqueous humour, the right eye of each animal is enucleated and dissected into front (anterior) and back (posterior) sections around the lens. Each section is weighed and homogenised in 500 μL of sterile PBS followed by 20 minutes centrifugation at 12000 rpm at 4° C. The resulting supernatant is divided into 3 aliquots and stored at −80° C. until subsequent cytokine analysis by R&D DuoSet ELISA.

Summary of In Vitro and In Vivo Screening Results

The compound of Example 2(k) is substantially more potent than the Reference Compound, displaying lower dissociation constants in the biochemical JAK family enzyme binding assays carried out at DiscoveRx (Table 1).

TABLE 1 Dissociation constants for selected kinases determined by LeadHunter Discover Services (DiscoveRx Corporation, Fremont, CA), using the KINOMEscan ™ technology. Dissociation Constant (nM) Test compound JAK1 JAK2 JAK3 TYK2 Reference Compound 2.7 0.78 1.9 1.9 Example 2(k) 1.0 0.047 0.092 0.13

Similarly, compounds of the examples of the present invention may be substantially more potent than the Reference Compound on inhibiting IFNγ release from CD3/IL2-stimulated PBMC cells (assay (a) above) and/or may display enhanced viabilities in cell cytotoxicity assay (c) above (Table 2).

TABLE 2 Effects of the compounds of the invention on inhibition of IFNγ release from stimulated PBMC cells (assay (a) above) and cell viability in Jurkat cells (assay (c) above). Test compound PBMC IFNγ IC₅₀ (nM) % viability at 1 μg/mL Reference Compound 81.3 49 Example 1 22.6 14 Example 2(a) 731.5 — Example 2(b) 87.0 — Example 2(c) 37.0 — Example 2(d) 269.9 — Example 2(e) 57.6 96 Example 2(f) 49.3 16 Example 2(g) 270.8 — Example 2(h) 12.1 56 Example 2(i) 14.5 53 Example 2(j) 5.3 12 Example 2(k) 9.9 93 Example 2(l) 9.4 10 Example 2(m) 15.2 88 Example 2(n) 29.2 87 Example 2(o) 22.5 62 Example 2(p) 153.7 — Example 2(q) 113.2 94 Example 2(r) 90.8 — Example 2(s) 693.6 — Example 2(t) 15.2 18 Example 2(u) 73.8 — Example 2(v) 7.5 82 Example 2(w) 941.5 —

Summary of Additional Studies

Determination of Solubilities in Fasted-State Simulated Colonic Fluid (FaSSCoF)

The solubilities of compounds of the invention in FaSSCoF at pH 6.5 are determined using a modification of a previously-reported procedure (Vertzoni, M., et al. Pharm. Res. 2010, 27, 2187-2196). In place of the bile salt extract employed in the original procedure (which extract is no longer available), the modified procedure uses a mixture of sodium taurochlorate (0.15 g), glycocholic acid (0.15 g), ursodeoxycholic acid (0.05 g), cholic acid (0.05 g), and glycodeoxycholic acid (0.05 g). These five bile acids are ground together with a mortar and pestle to produce a fine white powder that is incorporated into the FaSSCoF, as outlined below.

FaSSCoF Medium:

Tris(hydroxymethyl)aminomethane (Tris; 0.275 g) and maleic acid (0.44 g) are dissolved in water (35 mL) to give a solution whose pH is adjusted to 6.5 by treatment with 0.5M NaOH (ca. 12 mL). The solution is then made up to 50 mL with water. A portion of this Tris/maleate buffer solution (ca. 25 mL) is added to a 0.5 L round-bottomed flask, before being treated with 0.00565 g of the bile acid mixture described above. Solutions of phosphatidylcholine (0.0111 g) in DCM (0.15 mL) and palmitic acid (0.0013 g) in DCM (0.15 mL) are added, then the organic solvent is evaporated off under reduced pressure at 40° C. until a clear solution, with no perceptible DCM odour, is achieved. The volume of the evaporated solution is adjusted to 50 mL by addition of the remainder of Tris/maleate buffer, then BSA (0.115 g) is added, before being dissolved by gentle agitation.

Solubility Determination:

Test compounds are suspended in the pH 6.5 FaSSCoF medium to give a maximum final concentration of 2-10 mg/mL. The suspensions are equilibrated at 25° C. for 24 h, before being filtered through a glass fibre C filter. The filtrates are then diluted as appropriate for injection and quantification by HPLC with reference to a standard. Different volumes of the standard, diluted and undiluted sample solutions are injected and the solubilities are calculated using the peak areas determined by integration of the peak found at the same retention time as the principal peak in the standard injection.

FaSSCoF solubilities are shown in Table 3 below, which reveals that, unlike the Reference Compound, compounds of the Examples exhibited solubilities in the FaSSCoF medium at pH 6.5 of 0.01 mg/mL or greater.

TABLE 3 Solubilities measured for certain compounds of the Examples of the present invention in FaSSCoF at pH 6.5. pH 6.5 FaSSCoF Solubility (mg/mL) Test compound Run 1 Run 2 Reference Compound 0.006 0.006 Example 1 0.010 0.010 Example 2(h) 0.030 0.040 Example 2(k) 0.010 —

Determination of Pharmacokinetic Parameters

Studies were conducted by Sai Life Sciences (Hinjewadi, Pune, India) to investigate the systemic pharmacokinetics and total colon tissue distribution of compounds of the invention following a single oral administration in male C57BL/6 mice.

A group of twenty four male mice were administered with a solution of test compound formulated at a dose of 10 mg/10 mL/kg in aqueous hydroxypropyl-β-cyclodextrin (20% w/v in water). Blood samples (approximately 60 μL) were collected from retro-orbital plexus of each mouse under light isoflurane anaesthesia such that the samples were obtained at 0.5, 1, 2, 4, 6, 8, 12 and 24 hr. The blood samples were collected from a set of three mice at each time point in a labeled micro centrifuge tube containing K₂EDTA as anticoagulant. Plasma samples were separated by centrifugation at 4000 rpm for 10 min and stored below −70° C. until bioanalysis. After collection of blood sample, animals were humanely euthanized by carbon dioxide asphyxiation to collect total colon tissues. The colons were flushed with cold PBS (pH7.4) to remove contents. The total colon tissues were homogenized with cold PBS (pH7.4) of twice the weight of colon tissue and stored below −70° C. Total volume was three times the total colon tissue weights. All samples were processed for analysis by protein precipitation using acetonitrile and analyzed with a fit-for-purpose LC-MS/MS method. Pharmacokinetic parameters were calculated using the non-compartmental analysis tool of Phoenix WinNonlin® (Version 6.3).

The data catalogued in Table 4 reveal that, following oral administration, the compound of Example 2(k) achieved substantial colonic concentrations, while, in contrast, systemic plasma exposure was very low. Moreover, Example 2(k) demonstrates much higher colon-to-plasma ratios compared to the Reference Compound, as revealed in Table 5. In both of Tables 4 and 5, “P” refers to plasma and “C” refers to total colon.

TABLE 4 Mean plasma concentrations (ng/mL) or total colon levels (ng/g) obtained following oral administration of the Reference Compound and Example 2(k) to mice at 10 mg/kg in solution in 20% aqueous hydroxypropyl-β-cyclodextrin (dose volume = 10 mL/kg). Time (h) Compound Matrix 0.5 1 2 4 6 8 12 24 Reference P 5.2 12.8 2.2 14.4 114 95.0 0.0 0.0 Compound C 344 102 2,366 2,972 201 2,555 3,229 16.6 Example 2(k) P 2.4 2.6 1.9 1.1 3.4 4.8 1.2 0.0 C 431 1,155 5,447 7,232 4,815 15,217 2,224 44.8

TABLE 5 Total colon to plasma ratios obtained following oral administration of solutions of the Reference Compound and Example 2(k) at 10 mg/kg to mice. C_(max) AUC_(0-24 h) (ng/g or ng/mL) C_(max) (h · ng/g or h · ng/mL) AUC_(0-24 h) Compound C P ratio C P ratio Reference 3,229 114 28.2 43,739 368 119 Compound Example 2(k) 15,217 4.8 3,170 97,062 20 4,853

Abbreviations

-   -   aq aqueous     -   5-ASA 5-aminosalicylic acid     -   ATP adenosine-5′-triphosphate     -   BID bis in die (twice-daily)     -   Boc tert-butoxycarbonyl     -   br broad     -   BSA bovine serum albumin     -   CD Crohn's disease     -   COPD chronic obstructive pulmonary disease     -   d doublet     -   b chemical shift     -   DCM dichloromethane     -   DIPEA diisopropylethylamine     -   DMEM Dulbecco's modified eagle medium     -   DMF N,N-dimethylformamide     -   DMSO dimethyl sulfoxide     -   EDTA ethylenediaminetetraacetic acid     -   ELISA enzyme-linked immunosorbent assay     -   (ES⁻) electrospray ionization, negative mode     -   (ES⁺) electrospray ionization, positive mode     -   Et ethyl     -   Et₃N triethylamine     -   EtOAc ethyl acetate     -   EtOH ethanol     -   FaSSCoF fasted-state simulated colonic fluid     -   FBS foetal bovine serum     -   FCS foetal calf serum     -   HATU 2-(1H-7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyl uronium         hexafluoro-phosphate     -   HBSS Hank's balanced salt solution     -   HPLC high performance liquid chromatography     -   HPMC hydroxypropylmethylcellulose     -   h, hr or hrs hour(s)     -   HRP horseradish peroxidise     -   IBD Inflammatory bowel disease     -   IFNγ interferon-γ     -   IL interleukin     -   IPA isopropyl alcohol     -   JAK Janus kinase     -   JNK c-Jun N-terminal kinase     -   LC liquid chromatography     -   LPMC lamina propria mononuclear cells     -   LPS lipopolysaccharide     -   m multiplet     -   (M+H)⁺ protonated molecular ion     -   (M−H)⁻ deprotonated molecular ion     -   Me methyl     -   MeCN acetonitrile     -   MeOH methanol     -   MHz megahertz     -   min or mins minute(s)     -   MMAD mass median aerodynamic diameter     -   MPO myeloperoxidase     -   MS mass spectrometry     -   m/z mass-to-charge ratio     -   NMP N-Methyl-2-pyrrolidone     -   NMR nuclear magnetic resonance (spectroscopy)     -   OD optical density     -   PBMC peripheral blood mononuclear cell     -   PBS phosphate buffered saline     -   Ph phenyl     -   PHA phytohaemagglutinin     -   PI propidium iodide     -   PMA phorbol myristate acetate     -   q quartet     -   rt or RT room temperature     -   RP HPLC reverse phase high performance liquid chromatography     -   rpm revolutions per minute     -   RPMI Roswell Park Memorial Institute     -   s singlet     -   sat or satd saturated     -   SCID severe combined immunodeficiency     -   SCX solid supported cation exchange (resin)     -   SDS sodium dodecyl sulfate     -   S_(N)Ar nucleophilic aromatic substitution     -   Syk Spleen tyrosine kinase     -   t triplet     -   ^(t)Bu tert-butyl     -   T3P 1-propanephosphonic acid cyclic anhydride     -   TEA triethylamine     -   THF tetrahydrofuran     -   TFA trifluoroacetic acid     -   TGFβ transforming growth factor beta     -   TNFα tumor necrosis factor alpha     -   Tris tris(hydroxymethyl)aminomethane     -   TYK2 tyrosine kinase 2     -   UC ulcerative colitis     -   UPLC ultra performance liquid chromatography     -   UV ultra-violet

Prefixes n-, s-, i-, t- and tert- have their usual meanings: normal, secondary, iso, and tertiary. 

1. A compound of formula I,

wherein: X represents halo; Ak represents C₁₋₆ alkyl optionally substituted by one or more fluorine atoms; s represents 1 or 2; L represents C(O), S(O)₂, CH₂ or a bond; Q represents a 6-membered heteroaromatic ring containing 1, 2 or 3 N atoms; R¹ represents R³; R² represents H, methyl or R³; or R¹ and R², together with the N-atom to which they are attached, form a 4- to 7-membered heterocyclic group that is fully saturated or partially unsaturated, which heterocyclic group contains one N atom (the atom to which R¹ and R² are attached) and wherein (a) the heterocyclic group contains one further heteroatom that is S and is optionally substituted by one or more substituents selected from halo, hydroxy, oxo, C₁₋₄ alkyl, C₁₋₄ alkoxy and C₁₋₄ hydroxyalkyl or (b) the heterocyclic group optionally contains one or more further heteroatoms selected from N, O and S, is substituted by one or more substituents selected from C₁₋₄ hydroxyalkyl, C₁₋₄ carboxyalkyl and —(C₁₋₄ alkylene)-NR^(a)R^(b), and is optionally further substituted by one or more substituents selected from halo, hydroxy, oxo, CO₂H, C₁₋₄ alkyl and C₁₋₄ alkoxy or (c) the heterocyclic group is fused to a fully saturated or partially unsaturated 5- or 6-membered heterocyclic ring, which heterocylic ring contains one or more heteroatoms selected from from N, O and S and is optionally substituted with one or more substituents selected from oxo and C₁₋₄ alkyl; R³ represents Het^(x) or C₁₋₆ alkyl substituted by CO₂H, —(OCH₂CH₂)₀₋₄OR^(c), Het^(y), Het^(z), NR^(d)C(O)R^(e) or S(O)₁₋₂R^(f), which C₁₋₆ alkyl group is optionally further substituted by one or more substituents selected from halo, hydroxy and oxo; R^(a), R^(b), R^(c), R^(d) and R^(e) independently represent H or C₁₋₄ alkyl, or R^(a) and R^(b), together with the N-atom to which they are attached, form a 4- to 7-membered heterocyclic group that is fully saturated, partially unsaturated or fully aromatic and which heterocyclic group contains one N atom (the atom to which R^(a) and R^(b) are attached) and, optionally, one or more further heteroatoms selected from O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from halo, hydroxy, oxo, C₁₋₄ alkyl, C₁₋₄ alkoxy and C₁₋₄ hydroxyalkyl; R^(f) represents C₁₋₆ alkyl or C₃₋₇ cycloalkyl; Het^(x) represents, independently upon each occurrence, a 4- to 7-membered heterocyclic group that is fully saturated, partially unsaturated or fully aromatic, which group contains one or more heteroatoms selected from N, O and S, and which group is optionally substituted by one or more substituents selected from C₁₋₄ alkyl, halo, hydroxy and oxo; Het^(y) represents, independently upon each occurrence, a 5- to 10-membered heterocyclic group that is fully aromatic, which group contains one or more heteroatoms selected from N, O and S, and which group is optionally substituted by one or more substituents selected from C₁₋₄ alkyl, halo, hydroxy and oxo; and Het^(z) represents, independently upon each occurrence, a 4- to 7-membered heterocyclic group that is fully saturated or partially unsaturated, which group contains one or more heteroatoms selected from N, O and S, and which group is substituted by oxo or hydroxy, and which group is optionally further substituted by one or more substituents selected from C₁₋₄ alkyl, halo, hydroxy and oxo, or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof.
 2. A compound as claimed in claim 1 that is a compound of formula Ia, Ib or Ic,

or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof, wherein X, Ak, L, Q, R¹ and R² are as defined in claim
 1. 3. A compound as claimed in claim 1, wherein X represents fluoro and/or Ak represents C₁₋₃ alkyl.
 4. A compound as claimed in claim 1, wherein L represents C(O) and/or Q represents pyrazinyl.
 5. A compound as claimed in claim 1, wherein: R¹ represents R³; R² represents H, methyl or R³; or R¹ and R², together with the N-atom to which they are attached, form a 5- or 6-membered heterocyclic group that is fully saturated, which heterocyclic group contains one N atom (the atom to which R¹ and R² are attached) and wherein either (a) the heterocyclic group contains one further heteroatom that is S and the heterocyclic group is optionally substituted by one or more substituents selected from hydroxy, oxo and methyl or (b) the heterocyclic group optionally contains one further heteroatom selected from N, O and S, is substituted by a substituent selected from C₁₋₂ hydroxyalkyl (e.g. hydroxymethyl) and —(C₁₋₃ alkylene)-NR^(a)R^(b), and is optionally further substituted by methyl.
 6. A compound as claimed in claim 1, wherein: R³ represents Het^(x) or C₁₋₄ alkyl substituted by CO₂H, —(OCH₂CH₂)₀₋₂OR^(c), Het^(y), Het^(z) or NR^(d)C(O)R^(e); and/or R^(d) and R^(e) independently represent H or methyl.
 7. A compound as claimed in claim 1, wherein: R^(a) and R^(b) represent H or methyl, or R^(a) and R^(b), together with the N-atom to which they are attached, form a 5- or 6-membered heterocyclic group that is fully aromatic and which heterocyclic group contains one N atom (the atom to which R^(a) and R^(b) are attached) and, optionally, one or two further heteroatoms selected from O, S and N, and which heterocyclic group is optionally substituted by one or more substituents selected from methyl, methoxy and hydroxymethyl; and/or R^(c) represents H or methyl.
 8. A compound as claimed in claim 1, wherein: Het^(x) represents, independently upon each occurrence, a 5- or 6-membered heterocyclic group that is fully saturated, which group contains one or two heteroatoms selected from N, O and S, and which group is optionally substituted by one or two substituents selected from methyl, hydroxy and oxo; Het^(y) represents, independently upon each occurrence, a 5- or 6-membered heterocyclic group that is fully aromatic, which group contains one or two heteroatoms selected from N, O and S and which group is optionally substituted by one or two methyl substituents; and/or Het^(z) represents, independently upon each occurrence, a 5- or 6-membered heterocyclic group that is fully saturated, which group contains one or two heteroatoms selected from N, O and S, and which group is substituted by oxo or hydroxy, and which group is optionally further substituted by one or more substituents selected from methyl, hydroxy and oxo.
 9. A compound as claimed in claim 1 which is a compound selected from the list comprising: (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(methyl(1-methylpiperidin-4-yl)amino)pyrazin-2-yl)methanone; [TOP4026] 4-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)butanoic acid; [TOP4025] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(4-hydroxypiperidin-1-yl)ethyl)amino)pyrazin-2-yl)methanone; [TOP4027] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(methyl(pyridin-2-ylmethyl)amino)pyrazin-2-yl)methanone; [TOP4028] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(1-oxidothiomorpholino)ethyl)amino)pyrazin-2-yl)methanone; [TOP4029] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(1-oxidothiomorpholino)pyrazin-2-yl)methanone; [TOP4030] (5-(4-(2-(dimethylamino)ethyl)piperazin-1-yl)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone; [TOP4031] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-((2-(4-hydroxy-1-methylpiperidin-4-yl)ethyl)amino)pyrazin-2-yl)methanone; [TOP4032] (S)-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)(5-(2-(hydroxymethyl)pyrrolidin-1-yl)pyrazin-2-yl)methanone; [TOP4033] (5-(bis(2-methoxyethyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone; [TOP4034] (5-(4-(2-(1H-imidazol-1-yl)ethyl)piperazin-1-yl)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-1,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridin-5-yl)methanone; [TOP4035] 1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)ethyl)pyrrolidin-2-one; [TOP4036] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-(4-(2-hydroxyethyl)piperazin-1-yl)pyrazin-2-yl)methanone; [TOP4046] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2-(2-(2-methoxyethoxy)ethoxy)ethyl)amino)pyrazin-2-yl)methanone; [TOP4047] N-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)ethyl)-N-methylacetamide; [TOP4048] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5 (4H)-yl)(5-(3-(hydroxymethyl)azetidin-1-yl)pyrazin-2-yl)methanone; [TOP4049] (5-(bis(2-hydroxyethyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5 (4H)-yl)methanone; [TOP4059] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5 (4H)-yl)(5-((2-(methylsulfonyl)ethyl)amino)pyrazin-2-yl)methanone; [TOP4060] 1-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)amino)ethyl)pyrrolidin-2-one; [TOP4062] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((2R,3R,4R,5S)-3,4,5-trihydroxy-2-(hydroxymethyl)piperidin-1-yl)pyrazin-2-yl)methanone; [TOP4066] (2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)(5-((3aR,6aS)-5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)pyrazin-2-yl)methanone; [TOP4067] 3-(2-((5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)(methyl)amino)ethyl)oxazolidin-2-one; [TOP4068] (5-((2-(1,1-dioxidoisothiazolidin-2-yl)ethyl)(methyl)amino)pyrazin-2-yl)(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-6,7-dihydro-1H-imidazo[4,5-c]pyridin-5(4H)-yl)methanone; and [TOP4069] (3aR,6aS)-5-(5-(2-(6-(2-ethyl-5-fluoro-4-hydroxyphenyl)-1H-indazol-3-yl)-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-5-carbonyl)pyrazin-2-yl)tetrahydropyrrolo[3,4-c]pyrrole-1,3(2H,3aH)-dione, [TOP4071] or a pharmaceutically acceptable salt, solvate or isotopic derivative thereof.
 10. A pharmaceutical formulation comprising a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier.
 11. A combination product comprising (A) a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, and (B) another therapeutic agent, wherein each of components (A) and (B) is formulated in admixture with a pharmaceutically-acceptable adjuvant, diluent or carrier. 12-16. (canceled)
 17. A method of treating or preventing an inflammatory disease, said method comprising administering to a subject an effective amount of a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, or a pharmaceutical formulation comprising a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, in admixture with a pharmaceutically acceptable adjuvant, diluent or carrier, or a combination product comprising (A) a compound as defined in claim 1, or pharmaceutically acceptable salt, solvate or isotopic derivative thereof, and (B) another therapeutic agent, wherein each of components (A) and (B) is formulated in admixture with a pharmaceutically-acceptable adjuvant, diluent or carrier.
 18. A process for the preparation of a compound of formula I, as defined in claim 1, which process comprises: (a) reaction of a compound of formula II,

wherein X, Ak, s, L and Q are as defined in claim 1 and LG represents a leaving group, with a compound of formula III,

wherein R¹ and R² are as defined in claim 1; (b) for compounds of formula I in which L represents C(O), reaction of a compound of formula IV,

wherein X, Ak and s as defined in claim 1, with a compound of formula VIII,

wherein Q, R¹ and R² are as defined in claim 1; (c) reaction of a compound of formula IV, as defined above, with a compound of formula IX,

wherein LG¹ represents a leaving group and L, Q, R¹ and R² are as defined in claim 1; (d) for compounds of formula I in which L represents CH₂, reductive alkylation of a compound of formula IV, as defined above, with a compound of formula X,

wherein Q, R¹ and R² are as defined in claim 1, in the presence of a reducing agent; or (e) deprotection of a protected derivative of a compound of formula I wherein the protected derivative bears a protecting group on an O- or N-atom of the compound of formula I.
 19. A method according to claim 17, wherein the inflammatory disease is selected from the list comprising cystic fibrosis, pulmonary hypertension, lung sarcoidosis, idiopathic pulmonary fibrosis, COPD, chronic bronchitis, emphysema, asthma, paediatric asthma, atopic dermatitis, allergic dermatitis, contact dermatitis or psoriasis, allergic rhinitis, rhinitis, sinusitis, conjunctivitis, allergic conjunctivitis, keratoconjunctivitis sicca, dry eye, xerophthalmia, glaucoma, diabetic retinopathy, macular oedema, diabetic macular oedema, central retinal vein occlusion (CRVO), dry and/or wet age related macular degeneration (AMD), post-operative cataract inflammation, uveitis (including posterior, anterior and pan uveitis), corneal graft and limbal cell transplant rejection, gluten sensitive enteropathy (coeliac disease), eosinophilic esophagitis, intestinal graft versus host disease, Crohn's disease and ulcerative colitis.
 20. A method according to claim 17, wherein the inflammatory disease is uveitis, keratoconjunctivitis sicca, dry eye, xerophthalmia, Crohn's disease, ulcerative colitis, asthma or COPD. 